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Volume 30,Issue 7,2023 Table of Contents
2023, 30(7):541-551.
DOI: 10.3872/j.issn.1007-385X.2023.07.001
Abstract:
In recent years, with the in-depth understanding of tumor antigen and mechanism of antitumor immune response, therapeutic cancer vaccines have developed rapidly and are expected to be an important tool for clinical cancer therapy. Meanwhile, breakthroughs in key technologies, such as neo-antigen screening, vaccine design, vaccine delivery system and adjuvant, have further accelerated the development in this field. As for product research and development, many international pharmaceutical companies and emerging biotech companies are laying out various projects of therapeutic cancer vaccines, and several products have been approved for marketing, but the clinical outcomes are not ideal. Although the majority of current therapeutic cancer vaccines are in preclinical and clinical stages, they demonstrate good application prospect and market value. Present review provides an overview of therapeutic cancer vaccines from the perspective of research and development at home and abroad, mainly focusing on the currently fast-growing personalized neo-antigen vaccine, DC-based vaccine and mRNA vaccine. Meanwhile, it also summarizes the current challenges and envisions its to future development, which may provide some clues for research and product development of therapeutic cancer vaccines.
In recent years, with the in-depth understanding of tumor antigen and mechanism of antitumor immune response, therapeutic cancer vaccines have developed rapidly and are expected to be an important tool for clinical cancer therapy. Meanwhile, breakthroughs in key technologies, such as neo-antigen screening, vaccine design, vaccine delivery system and adjuvant, have further accelerated the development in this field. As for product research and development, many international pharmaceutical companies and emerging biotech companies are laying out various projects of therapeutic cancer vaccines, and several products have been approved for marketing, but the clinical outcomes are not ideal. Although the majority of current therapeutic cancer vaccines are in preclinical and clinical stages, they demonstrate good application prospect and market value. Present review provides an overview of therapeutic cancer vaccines from the perspective of research and development at home and abroad, mainly focusing on the currently fast-growing personalized neo-antigen vaccine, DC-based vaccine and mRNA vaccine. Meanwhile, it also summarizes the current challenges and envisions its to future development, which may provide some clues for research and product development of therapeutic cancer vaccines.
2023, 30(7):552-559.
DOI: 10.3872/j.issn.1007-385X.2023.07.002
Abstract:
Objective: To investigate the effect of miR-216b-5p on cisplatin (DDP) resistance in esophageal cancer Eca109 cells and its mechanism. Methods: The expression levels of miR-216b-5p in esophageal cancer cells TE-1, KYSE-150, Eca109 and drug resistant Eca109/DDP cells were detected by qPCR. The miR-216b-5p mimics, mimic NC and autophagy related protein 5 (ATG5) over-expressed plasmids were transfected into Eca109/DDP cells by liposome transfection technique. The proliferation and apoptosis of transfected Eca109/DDP cells were detected by CCK-8, EdU methods and flow cytometry, respectively. The occurrence of autophagy in each group of cells after transfection with mRFP-eGFP-LC3 lentivirus was examined by mRFP-eGFP-LC3 dual fluorescent labeling assay, and the expression of autophagy-related markers, LC3, Beclin 1 and P62, was detected by WB. The targeting relationship between miR-216b-5p and ATG5 was verified by luciferase reporter gene assay, and the expression of ATG5 was detected by Western blotting. A nude mouse Eca109/DDP cell transplanted tumor model was established to observe the effect of miR-216b-5p over-expression on the growth of transplanted tumor. Results: miR-216b-5p was lowly expressed in TE-1, KYSE-150, Eca109 and Eca109/DDP cells (all P<0.05). Over-expression of miR-216b-5p significantly inhibited the proliferation and induced apoptosis of Eca109/DDP cells (both P<0.05), reduced the number of autophagosomes in transfected cells (P<0.05), and down-regulated LC3Ⅱ/LC3Ⅰratio and Beclin 1 protein level, but up-regulated P62 protein level (all P<0.05). Luciferase reporter gene assay confirmed that miR-216b-5p negatively regulated ATG5 expression (P<0.05). Over-expression of ATG5 could significantly weaken the effects of miR-216b-5p mimic on suppressing proliferation, autophagy and inducing apoptosis of Eca109/DDP cells (all P<0.05), and reduce the expression of autophagy related protein P62, but increase the LC3Ⅱ/LC3Ⅰratio and Beclin 1 protein level (all P<0.05). The tumor bearing experiments showed that over-expression of miR-216b-5p could significantly inhibit the growth of transplanted tumors in nude mice (P<0.05).Conclusion: Over-expression of miR-216b-5p can reverse DDP resistance in Eca109/DDP cells, and the mechanism may be related to its negative regulation of autophagy-related gene ATG5 to affect cell autophagy.
Objective: To investigate the effect of miR-216b-5p on cisplatin (DDP) resistance in esophageal cancer Eca109 cells and its mechanism. Methods: The expression levels of miR-216b-5p in esophageal cancer cells TE-1, KYSE-150, Eca109 and drug resistant Eca109/DDP cells were detected by qPCR. The miR-216b-5p mimics, mimic NC and autophagy related protein 5 (ATG5) over-expressed plasmids were transfected into Eca109/DDP cells by liposome transfection technique. The proliferation and apoptosis of transfected Eca109/DDP cells were detected by CCK-8, EdU methods and flow cytometry, respectively. The occurrence of autophagy in each group of cells after transfection with mRFP-eGFP-LC3 lentivirus was examined by mRFP-eGFP-LC3 dual fluorescent labeling assay, and the expression of autophagy-related markers, LC3, Beclin 1 and P62, was detected by WB. The targeting relationship between miR-216b-5p and ATG5 was verified by luciferase reporter gene assay, and the expression of ATG5 was detected by Western blotting. A nude mouse Eca109/DDP cell transplanted tumor model was established to observe the effect of miR-216b-5p over-expression on the growth of transplanted tumor. Results: miR-216b-5p was lowly expressed in TE-1, KYSE-150, Eca109 and Eca109/DDP cells (all P<0.05). Over-expression of miR-216b-5p significantly inhibited the proliferation and induced apoptosis of Eca109/DDP cells (both P<0.05), reduced the number of autophagosomes in transfected cells (P<0.05), and down-regulated LC3Ⅱ/LC3Ⅰratio and Beclin 1 protein level, but up-regulated P62 protein level (all P<0.05). Luciferase reporter gene assay confirmed that miR-216b-5p negatively regulated ATG5 expression (P<0.05). Over-expression of ATG5 could significantly weaken the effects of miR-216b-5p mimic on suppressing proliferation, autophagy and inducing apoptosis of Eca109/DDP cells (all P<0.05), and reduce the expression of autophagy related protein P62, but increase the LC3Ⅱ/LC3Ⅰratio and Beclin 1 protein level (all P<0.05). The tumor bearing experiments showed that over-expression of miR-216b-5p could significantly inhibit the growth of transplanted tumors in nude mice (P<0.05).Conclusion: Over-expression of miR-216b-5p can reverse DDP resistance in Eca109/DDP cells, and the mechanism may be related to its negative regulation of autophagy-related gene ATG5 to affect cell autophagy.
2023, 30(7):560-567.
DOI: 10.3872/j.issn.1007-385X.2023.07.003
Abstract:
Objective: To explore the effects of brucea javanica oil emulsion (BJOE) on proliferation, apoptosis and autophagy of esophageal squamous cell carcinoma TE-1 cells and the possible mechanism. Methods: According to the different interventions TE-1 cells were divided into control group, RAPA (autophagy agonist) group, 740Y-P (PI3K activator) group, BJOE group, BJOE+RAPA group and BJOE+740Y-P group. Cell apoptosis, proliferation, migration and invasion were detected by FCM, clonogenesis and Transwell assay; the mRNA expression of PI3K, Akt, mTOR, LC3Ⅰ, LC3Ⅱ, p62, Beclin 1 and caspase-3 in cells was detected by qPCR; and the protein expression levels of PI3K,Akt, mTOR and their phosphorylation as well as the protein expression of LC3Ⅱ/Ⅰ, p62, Beclin1 and caspase-3 were detected by Western blotting. Results: Compared with the control group, RAPA group and BJOE group exhibited increased cell apoptosis rate (both P<0.01),reduced clone formation rate, cell invasion and migration ability (all P<0.01), decreased mRNA and protein phosphorylation levels of PI3K,Akt, mTOR (all P<0.05) as well as decreased mRNA and protein levels of p62 (all P<0.01), and increased mRNA and protein levels of LC3 Ⅱ/Ⅰ, Beclin 1 and caspase-3 (all P<0.05); however, the results of 740Y-P group were opposite (all P<0.05). Compared with RAPA group or 740Y-P group alone, BJOE+RAPA or BJOE+740Y-P group exhibited increased apoptosis rate (P<0.01), decreased clone formation rate,cell invasion and migration ability (all P<0.01), decreased mRNA and protein phosphorylation levels of PI3K, Akt, mTOR (all P<0.05) as well as decreased mRNA and protein levels of p62 (all P<0.05), and increased mRNA and protein levels of LC3Ⅱ/Ⅰ, Beclin 1 and caspase-3 (all P<0.05). Conclusion: BJOE significantly inhibits the proliferation, invasion, migration and promotes apoptosis and autophagy of TE-1 cells, and the mechanism may be related to inhibition of the activation of PI3K/Akt/mTOR signaling pathway.
Objective: To explore the effects of brucea javanica oil emulsion (BJOE) on proliferation, apoptosis and autophagy of esophageal squamous cell carcinoma TE-1 cells and the possible mechanism. Methods: According to the different interventions TE-1 cells were divided into control group, RAPA (autophagy agonist) group, 740Y-P (PI3K activator) group, BJOE group, BJOE+RAPA group and BJOE+740Y-P group. Cell apoptosis, proliferation, migration and invasion were detected by FCM, clonogenesis and Transwell assay; the mRNA expression of PI3K, Akt, mTOR, LC3Ⅰ, LC3Ⅱ, p62, Beclin 1 and caspase-3 in cells was detected by qPCR; and the protein expression levels of PI3K,Akt, mTOR and their phosphorylation as well as the protein expression of LC3Ⅱ/Ⅰ, p62, Beclin1 and caspase-3 were detected by Western blotting. Results: Compared with the control group, RAPA group and BJOE group exhibited increased cell apoptosis rate (both P<0.01),reduced clone formation rate, cell invasion and migration ability (all P<0.01), decreased mRNA and protein phosphorylation levels of PI3K,Akt, mTOR (all P<0.05) as well as decreased mRNA and protein levels of p62 (all P<0.01), and increased mRNA and protein levels of LC3 Ⅱ/Ⅰ, Beclin 1 and caspase-3 (all P<0.05); however, the results of 740Y-P group were opposite (all P<0.05). Compared with RAPA group or 740Y-P group alone, BJOE+RAPA or BJOE+740Y-P group exhibited increased apoptosis rate (P<0.01), decreased clone formation rate,cell invasion and migration ability (all P<0.01), decreased mRNA and protein phosphorylation levels of PI3K, Akt, mTOR (all P<0.05) as well as decreased mRNA and protein levels of p62 (all P<0.05), and increased mRNA and protein levels of LC3Ⅱ/Ⅰ, Beclin 1 and caspase-3 (all P<0.05). Conclusion: BJOE significantly inhibits the proliferation, invasion, migration and promotes apoptosis and autophagy of TE-1 cells, and the mechanism may be related to inhibition of the activation of PI3K/Akt/mTOR signaling pathway.
2023, 30(7):568-576.
DOI: 10.3872/j.issn.1007-385X.2023.07.004
Abstract:
Objective: To investigate the effect and mechanism of live attenuated measles vaccine strain 191 (MV-Hu191) on triple negative breast cancer MDA-MB-231 and 4T1 cells in vitro and in vivo based on proteomics. Methods: CCK-8 method was used to analyze the effect of MV-Hu191 on the proliferation of MDA-MB-231 and 4T1 cells. Liquid chromatography-mass spectrometry was used to analyze the effect of MV-Hu191 treatment on protein spectrum in MDA-MB-231 cells. Multiple databases were used to screen typical differentially expressed proteins, followed with GO, KEGG, subcellular localization and functional annotation. Intratumoral injection of 1×106 TCID50 MV-Hu191 was used to intervene the growth of 4T1 cell transplanted tumor in mice. Flow cytometry was applied to detect T cell subpopulations in splenic tissue, and ELISA was used for the detection of serum levels of TNF-α and IL-6. Results: In vitro experiments showed that MV-Hu191 could significantly inhibit the proliferation of MDA-MB-231 and 4T1 cells (P<0.01). Proteomic analysis showed that 38 proteins were significantly upregulated while 12 proteins were downregulated in MDA-MB-231 cells after MV-Hu191 treatment. The differentially expressed proteins were mainly involved in the biological processes of cell adhesion, signaling receptor activation, cell metabolism, and stress responses. The subcellular localization of 22 differentially expressed proteins was located outside the cell. The KEGG functional classification showed that the most differentially expressed proteins were related to immunomodulatory functions and they were all upregulated, including C4A, C8B, SERPINF2, A2M, SERPINC1, CTSB, SERPING1, and C5; PPI prediction found immune-related differential proteins were associated with CD4, CD8, TNF-ɑ and IL-6. In vivo experiments showed that the number of CD4+ T cells in spleen tissues of mice in the MV-Hu191 intervention group was higher than that in the control group, but the difference was not significant (P>0.05). The ratio of CD4+/CD8+ T cells and the serum levels of TNF-ɑ and IL-6 in the MV-Hu191 intervention group were significantly higher than those in the control group (P<0.05, P<0.01). Conclusion: MV-Hu191 significantly inhibits the proliferation of MDA-MB-231 and 4T1 cells and antagonizes the tumorigenicity in mice bearing 4T1 cell xenograft. The mechanism may be that MV-Hu191 achieve its anti-tumor effect by activating immune effector molecules.
Objective: To investigate the effect and mechanism of live attenuated measles vaccine strain 191 (MV-Hu191) on triple negative breast cancer MDA-MB-231 and 4T1 cells in vitro and in vivo based on proteomics. Methods: CCK-8 method was used to analyze the effect of MV-Hu191 on the proliferation of MDA-MB-231 and 4T1 cells. Liquid chromatography-mass spectrometry was used to analyze the effect of MV-Hu191 treatment on protein spectrum in MDA-MB-231 cells. Multiple databases were used to screen typical differentially expressed proteins, followed with GO, KEGG, subcellular localization and functional annotation. Intratumoral injection of 1×106 TCID50 MV-Hu191 was used to intervene the growth of 4T1 cell transplanted tumor in mice. Flow cytometry was applied to detect T cell subpopulations in splenic tissue, and ELISA was used for the detection of serum levels of TNF-α and IL-6. Results: In vitro experiments showed that MV-Hu191 could significantly inhibit the proliferation of MDA-MB-231 and 4T1 cells (P<0.01). Proteomic analysis showed that 38 proteins were significantly upregulated while 12 proteins were downregulated in MDA-MB-231 cells after MV-Hu191 treatment. The differentially expressed proteins were mainly involved in the biological processes of cell adhesion, signaling receptor activation, cell metabolism, and stress responses. The subcellular localization of 22 differentially expressed proteins was located outside the cell. The KEGG functional classification showed that the most differentially expressed proteins were related to immunomodulatory functions and they were all upregulated, including C4A, C8B, SERPINF2, A2M, SERPINC1, CTSB, SERPING1, and C5; PPI prediction found immune-related differential proteins were associated with CD4, CD8, TNF-ɑ and IL-6. In vivo experiments showed that the number of CD4+ T cells in spleen tissues of mice in the MV-Hu191 intervention group was higher than that in the control group, but the difference was not significant (P>0.05). The ratio of CD4+/CD8+ T cells and the serum levels of TNF-ɑ and IL-6 in the MV-Hu191 intervention group were significantly higher than those in the control group (P<0.05, P<0.01). Conclusion: MV-Hu191 significantly inhibits the proliferation of MDA-MB-231 and 4T1 cells and antagonizes the tumorigenicity in mice bearing 4T1 cell xenograft. The mechanism may be that MV-Hu191 achieve its anti-tumor effect by activating immune effector molecules.
2023, 30(7):577-585.
DOI: 10.3872/j.issn.1007-385X.2023.07.005
Abstract:
Objective: To investigate the effects of DJ-1 gene over-expression on proliferation, migration, invasion and epithelial-mesenchymal transformation (EMT) of human gastric cancer MGC803 cells and the underlying mechanism. Methods: MGC803 cells with DJ-1 over-expression were constructed by gene transfection technology. Three groups of cells, namely MGC803 group, empty vector group, and DJ-1 over-expression group were set. The effects of DJ-1 gene over-expression on proliferation, clone formation, migration and invasion of MGC803 cells were detected by MTT, plate cloning assay, cell scratch assay and Transwell invasion assay, respectively. The effects of DJ-1 over-expression on the expression levels of DJ-1, PTEN, Akt, p-Akt, Snail, vimentin, E-cadherin, MMP-9 and TIMP-3 were detected by qPCR and Western blot. The morphological changes of MGC803 cells were observed by phase contrast microscope. The effect of DJ-1 over-expression on the growth of MGC803 cell transplanted tumor in vivo was detected in nude mice. Results: MGC803 cells with stable DJ-1 over-expression were constructed successfully. The proliferation ability and number of clones in DJ-1 over-expression group were significantly increased compared with those in MGC803 cell group and empty vector group (all P<0.05); the cell migration distance was significantly increased while the scratch distance was significantly shortened in DJ-1 over-expression group compared with those in MGC803 cell group and empty vector group (all P<0.05); and the migrated and invaded cells of DJ-1 over-expression group were significantly more than those of MGC803 group and empty vector group (all P<0.05). Moreover, the expression of DJ-1 was significantly up-regulated while the expression of PTEN was significantly down-regulated at both the mRNA and protein levels in the DJ-1 over-expression group compared with those in MGC803 group and empty vector group (both P<0.05). There was no significant difference in total Akt protein among all groups (P>0.05), but the expression of p-Akt protein in DJ-1 over-expression group was significantly up-regulated compared with MGC803 group and empty vector group (all P<0.05). In addition, Snail, vimentin and MMP-9 were up-regulated in DJ-1 over-expression group, while E-cadherin and TIMP-3 were down-regulated (all P<0.05). Phase contrast microscopy showed that the number of long spindle cells increased while the number of round and oval cells decreased, and the atypia was more obvious in the DJ-1 over-expression group. In vivo experiments showed that the growth rate of transplanted tumor in DJ-1 over-expression group was significantly accelerated, and the weight of transplanted tumor was significantly increased (both P<0.05) as compared with MGC803 group. Conclusion: DJ-1 over-expression can inhibit the proliferation, migration, invasion and EMT of MGC803 cells both in vitro and in vivo through PTEN/Akt pathway.
Objective: To investigate the effects of DJ-1 gene over-expression on proliferation, migration, invasion and epithelial-mesenchymal transformation (EMT) of human gastric cancer MGC803 cells and the underlying mechanism. Methods: MGC803 cells with DJ-1 over-expression were constructed by gene transfection technology. Three groups of cells, namely MGC803 group, empty vector group, and DJ-1 over-expression group were set. The effects of DJ-1 gene over-expression on proliferation, clone formation, migration and invasion of MGC803 cells were detected by MTT, plate cloning assay, cell scratch assay and Transwell invasion assay, respectively. The effects of DJ-1 over-expression on the expression levels of DJ-1, PTEN, Akt, p-Akt, Snail, vimentin, E-cadherin, MMP-9 and TIMP-3 were detected by qPCR and Western blot. The morphological changes of MGC803 cells were observed by phase contrast microscope. The effect of DJ-1 over-expression on the growth of MGC803 cell transplanted tumor in vivo was detected in nude mice. Results: MGC803 cells with stable DJ-1 over-expression were constructed successfully. The proliferation ability and number of clones in DJ-1 over-expression group were significantly increased compared with those in MGC803 cell group and empty vector group (all P<0.05); the cell migration distance was significantly increased while the scratch distance was significantly shortened in DJ-1 over-expression group compared with those in MGC803 cell group and empty vector group (all P<0.05); and the migrated and invaded cells of DJ-1 over-expression group were significantly more than those of MGC803 group and empty vector group (all P<0.05). Moreover, the expression of DJ-1 was significantly up-regulated while the expression of PTEN was significantly down-regulated at both the mRNA and protein levels in the DJ-1 over-expression group compared with those in MGC803 group and empty vector group (both P<0.05). There was no significant difference in total Akt protein among all groups (P>0.05), but the expression of p-Akt protein in DJ-1 over-expression group was significantly up-regulated compared with MGC803 group and empty vector group (all P<0.05). In addition, Snail, vimentin and MMP-9 were up-regulated in DJ-1 over-expression group, while E-cadherin and TIMP-3 were down-regulated (all P<0.05). Phase contrast microscopy showed that the number of long spindle cells increased while the number of round and oval cells decreased, and the atypia was more obvious in the DJ-1 over-expression group. In vivo experiments showed that the growth rate of transplanted tumor in DJ-1 over-expression group was significantly accelerated, and the weight of transplanted tumor was significantly increased (both P<0.05) as compared with MGC803 group. Conclusion: DJ-1 over-expression can inhibit the proliferation, migration, invasion and EMT of MGC803 cells both in vitro and in vivo through PTEN/Akt pathway.
2023, 30(7):586-593.
DOI: 10.3872/j.issn.1007-385X.2023.07.006
Abstract:
Objective: To investigate the expression of circSMRCA5 in non-small cell lung cancer (NSCLC) tissues and cells and its potential function and mechanism in the occurrence and development of NSCLC. Methods: The expression of circSMARCA5 in NSCLC tissues was detected by qPCR. The circSMARCA5 over-expression plasmid and the control plasmid pLC5 were transfected into human lung cancer A549 and H1975 cells using the lentiviral transfection method, and the expression levels of circSMARCA5 in the stably transfected cell lines were detected by qPCR. The effects of circSMARCA5 over-expression on the biological behaviors of A549 and H1975 cells were examined by CCK-8, clone formation, cell cycle, and tumor xenografts. Mechanistically, transcriptome sequencing, KEGG and GO enrichment analysis were conducted to identify possible target genes of circSMARCA5. Subcutaneous xenograft models were constructed in BABL/c nude mice and immunocompetent C57 mice by using circSMARCA5-over-expressed NSCLC cells or Lewis cells. The effect of circSMARCA5 on the growth of subcutaneously transplanted tumors in nude mice was observed, and its effect on the contents of Treg cells in Lewis cells transplanted tumor tissues was detected by flow cytometry. Results: circSMARCA5 was highly expressed in NSCLC tissues (P<0.01). Over-expression of circSMARCA5 could promote the proliferation of NSCLC cells in vitro (P<0.05, P<0.01). In in vivo experiments,circSMARCA5 could promote the growth of the transplanted tumors in nude mice (P<0.01). Mechanistically, C-C chemokine ligand 5 (CCL5) was identified as a downstream target gene of circSMARCA5 by KEGG and GO enrichment analysis. The expression of CCL5 was increased in A549 and H1975 cells of circSMARCA5 over-expression group (P<0.05). circSMARCA5-mediated upregulation of CCL5 promoted the growth of subcutaneous tumors in immunocompetent C57 mice. Flow cytometry of single-cell suspensions prepared from subcutaneous tumors of C57 mice showed that the proportion of Treg cells in the circSMARCA5 over-expression group was higher than that in the control group ([3.1±0.5]% vs [1.0±0.1]%, P<0.05). Conclusion: circSMARCA5 is highly expressed in NSCLC tissues. circSMARCA5 may recruit Treg cells via CCL5, leading to the immune escape of tumors and promoting the progression of NSCLC.
Objective: To investigate the expression of circSMRCA5 in non-small cell lung cancer (NSCLC) tissues and cells and its potential function and mechanism in the occurrence and development of NSCLC. Methods: The expression of circSMARCA5 in NSCLC tissues was detected by qPCR. The circSMARCA5 over-expression plasmid and the control plasmid pLC5 were transfected into human lung cancer A549 and H1975 cells using the lentiviral transfection method, and the expression levels of circSMARCA5 in the stably transfected cell lines were detected by qPCR. The effects of circSMARCA5 over-expression on the biological behaviors of A549 and H1975 cells were examined by CCK-8, clone formation, cell cycle, and tumor xenografts. Mechanistically, transcriptome sequencing, KEGG and GO enrichment analysis were conducted to identify possible target genes of circSMARCA5. Subcutaneous xenograft models were constructed in BABL/c nude mice and immunocompetent C57 mice by using circSMARCA5-over-expressed NSCLC cells or Lewis cells. The effect of circSMARCA5 on the growth of subcutaneously transplanted tumors in nude mice was observed, and its effect on the contents of Treg cells in Lewis cells transplanted tumor tissues was detected by flow cytometry. Results: circSMARCA5 was highly expressed in NSCLC tissues (P<0.01). Over-expression of circSMARCA5 could promote the proliferation of NSCLC cells in vitro (P<0.05, P<0.01). In in vivo experiments,circSMARCA5 could promote the growth of the transplanted tumors in nude mice (P<0.01). Mechanistically, C-C chemokine ligand 5 (CCL5) was identified as a downstream target gene of circSMARCA5 by KEGG and GO enrichment analysis. The expression of CCL5 was increased in A549 and H1975 cells of circSMARCA5 over-expression group (P<0.05). circSMARCA5-mediated upregulation of CCL5 promoted the growth of subcutaneous tumors in immunocompetent C57 mice. Flow cytometry of single-cell suspensions prepared from subcutaneous tumors of C57 mice showed that the proportion of Treg cells in the circSMARCA5 over-expression group was higher than that in the control group ([3.1±0.5]% vs [1.0±0.1]%, P<0.05). Conclusion: circSMARCA5 is highly expressed in NSCLC tissues. circSMARCA5 may recruit Treg cells via CCL5, leading to the immune escape of tumors and promoting the progression of NSCLC.
2023, 30(7):594-602.
DOI: 10.3872/j.issn.1007-385X.2023.07.007
Abstract:
Objective: To investigate the expression of basic leucine zipper ATF-like transcription factor 3 (BATF3) in clear cell renal cell carcinoma (ccRCC) and the molecular mechanism of its regulation of malignant biological behavior of ccRCC cells. Methods: ccRCC tissues and para-cancerous tissues were collected from 64 ccRCC patients who underwent surgical treatment at the 980th Hospital of the Joint Logistics Support Force of PLA from March 2019 to January 2022. The expression of BATF3 mRNA in ccRCC tissues, para-cancerous tissues, renal carcinoma ACHN and 786-O cells was determined by qPCR. The expressions of BATF3 protein in ccRCC tissues and para-cancerous tissues were determined by immunohistochemistry and the relationship between the expressions and the clinicopathologic characteristics was analyzed. BATF3 knockdown and over-expression plasmids were constructed and transfected into 786-O and ACHN cells, and the effects of BATF3 on the proliferation, migration and invasion of 786-O cells or ACHN cells were determined by MTS and Transwell assays. The effects of BATF3 on the expressions of related epithelial-mesenchymal transition (EMT) genes of 786-O and ACHN cells were detected by qPCR. CHIP and dual luciferase reporter assay were performed to detect the binding of BATF3 as a transcription factor to vimentin (VIM) and regulate its transcription. The effects of simultaneous over-expression of BATF3 and knockdown of VIM on the proliferation, migration and invasion ability of 786-O cells were determined by MTS and Transwell assays. Results: Compared with para-cancerous tissues, BATF3 mRNA and protein expressions were markedly higher in ccRCC tissues (all P<0.01) and the expression level of BATF3 mRNA was closely correlated with the degree of differentiation of ccRCC and its TNM stage (all P<0.01). Compared with normal renal epithelial cells 293T, the expression of BATF3 was significantly higher in ccRCC cells ACHN and 786-O (all P<0.01). Knockdown of BATF3 expression significantly inhibited the proliferation,migration and invasion ability of 786-O cells (all P<0.01). Over-expression of BATF3 significantly promoted the proliferation,migration and invasion ability of ACHN cells (all P<0.01). Knockdown or overexpression of BATF3 inhibited the expression of EMT-related genes in 786-O cells or promoted the expression of EMT-related genes in ACHN cells (all P<0.01). BATF3 could bind to sites in the upstream promoter region of VIM and directly regulate the transcription expression of VIM. Simultaneous overexpression of BATF3 and knockdown of VIM reversed the effects of over-expression of BATF3 on the proliferation, migration and invasion ability of 786-O cells. Conclusion: BATF3 was highly expressed in ccRCC tissues and was closely related to its differentiation degree and TNM stage. BATF3 directly regulated the expression of VIM, which in turn regulated the malignant biological behavior of ACHN and 786-O cells. Therefore, it could be used as a potential target for clinical treatment of ccRCC.
Objective: To investigate the expression of basic leucine zipper ATF-like transcription factor 3 (BATF3) in clear cell renal cell carcinoma (ccRCC) and the molecular mechanism of its regulation of malignant biological behavior of ccRCC cells. Methods: ccRCC tissues and para-cancerous tissues were collected from 64 ccRCC patients who underwent surgical treatment at the 980th Hospital of the Joint Logistics Support Force of PLA from March 2019 to January 2022. The expression of BATF3 mRNA in ccRCC tissues, para-cancerous tissues, renal carcinoma ACHN and 786-O cells was determined by qPCR. The expressions of BATF3 protein in ccRCC tissues and para-cancerous tissues were determined by immunohistochemistry and the relationship between the expressions and the clinicopathologic characteristics was analyzed. BATF3 knockdown and over-expression plasmids were constructed and transfected into 786-O and ACHN cells, and the effects of BATF3 on the proliferation, migration and invasion of 786-O cells or ACHN cells were determined by MTS and Transwell assays. The effects of BATF3 on the expressions of related epithelial-mesenchymal transition (EMT) genes of 786-O and ACHN cells were detected by qPCR. CHIP and dual luciferase reporter assay were performed to detect the binding of BATF3 as a transcription factor to vimentin (VIM) and regulate its transcription. The effects of simultaneous over-expression of BATF3 and knockdown of VIM on the proliferation, migration and invasion ability of 786-O cells were determined by MTS and Transwell assays. Results: Compared with para-cancerous tissues, BATF3 mRNA and protein expressions were markedly higher in ccRCC tissues (all P<0.01) and the expression level of BATF3 mRNA was closely correlated with the degree of differentiation of ccRCC and its TNM stage (all P<0.01). Compared with normal renal epithelial cells 293T, the expression of BATF3 was significantly higher in ccRCC cells ACHN and 786-O (all P<0.01). Knockdown of BATF3 expression significantly inhibited the proliferation,migration and invasion ability of 786-O cells (all P<0.01). Over-expression of BATF3 significantly promoted the proliferation,migration and invasion ability of ACHN cells (all P<0.01). Knockdown or overexpression of BATF3 inhibited the expression of EMT-related genes in 786-O cells or promoted the expression of EMT-related genes in ACHN cells (all P<0.01). BATF3 could bind to sites in the upstream promoter region of VIM and directly regulate the transcription expression of VIM. Simultaneous overexpression of BATF3 and knockdown of VIM reversed the effects of over-expression of BATF3 on the proliferation, migration and invasion ability of 786-O cells. Conclusion: BATF3 was highly expressed in ccRCC tissues and was closely related to its differentiation degree and TNM stage. BATF3 directly regulated the expression of VIM, which in turn regulated the malignant biological behavior of ACHN and 786-O cells. Therefore, it could be used as a potential target for clinical treatment of ccRCC.
2023, 30(7):603-611.
DOI: 10.3872/j.issn.1007-385X.2023.07.008
Abstract:
Objective: To investigate the expression levels of high mobility group protein 1 (HMGB1) and indoleamine-2, 3-dioxygenase (IDO) in serum of patients with esophageal squamous cell carcinoma (ESCC) and their correlation with clinicopathologic features and lymphocyte subsets. Methods: Ninety-five ESCC patients initially hospitalized in the Fourth Hospital of Hebei Medical University from March 2021 to August 2022 were selected as the ESCC group, and the other 40 healthy subjects who underwent physical examination were selected as the control group. The serum levels of HMGB1 and IDO of all subjects and the contents of HMGB1, IDO and p65 in the supernatant of ESCC cell culture in different groups were detected by ELISA. The lymphocyte subsets in the peripheral blood of all subjects were detected by flow cytometry. WB was performed to determine the effect of HMGB1 knockdown as well as HMGB1 knockdown followed by addition of NF-κB signaling pathway activator on the expression levels of HMGB1, IDO and p65 in ESCC cells. Results:The serum levels of HMGB1 and IDO in ESCC group were significantly higher than those in control group (all P<0.01). Elevated serum HMGB1 and IDO levels were the independent risk factors for clinical progression of ESCC (all P<0.01), and their combined detection showed a higher predictive value for the clinical progression of ESCC (P<0.01). Serum HMGB1 and IDO levels were significantly correlated with T stage, N stage and clinical stage in ESCC patients (all P<0.05); Serum HMGB1 levels in ESCC group was negatively correlated with the absolute counts of CD3+ T cells, CD4+ T cells, B cells and NK cells in peripheral blood, and positively correlated with Treg cell percentage (all P<0.05); while serum IDO levels was negatively correlated with percentage and absolute count of CD3+ T cells, CD4+ T cells, and absolute counts of CD8+ T cells and B cells in peripheral blood,but positively correlated with the percentage of Treg cells (all P<0.05). Serum HMGB1 was positively correlated with IDO (P<0.01). The expression levels of IDO and p65 in KYSE30 and ECA109 cells and their culture supernatant in si-HMGB1 group were significantly lower than those in si-NC group and si-HMGB1+PMA group (all P<0.05). Conclusion: Serum HMGB1 and IDO are closely related to the clinical progression of ESCC and immune function of ESCC patients, and have the potential to serve as tumor markers and new immunotherapy targets of ESCC. HMGB1 may promote IDO expression through NF-κB signaling pathway, and dual-target combination therapy may achieve better efficacy.
Objective: To investigate the expression levels of high mobility group protein 1 (HMGB1) and indoleamine-2, 3-dioxygenase (IDO) in serum of patients with esophageal squamous cell carcinoma (ESCC) and their correlation with clinicopathologic features and lymphocyte subsets. Methods: Ninety-five ESCC patients initially hospitalized in the Fourth Hospital of Hebei Medical University from March 2021 to August 2022 were selected as the ESCC group, and the other 40 healthy subjects who underwent physical examination were selected as the control group. The serum levels of HMGB1 and IDO of all subjects and the contents of HMGB1, IDO and p65 in the supernatant of ESCC cell culture in different groups were detected by ELISA. The lymphocyte subsets in the peripheral blood of all subjects were detected by flow cytometry. WB was performed to determine the effect of HMGB1 knockdown as well as HMGB1 knockdown followed by addition of NF-κB signaling pathway activator on the expression levels of HMGB1, IDO and p65 in ESCC cells. Results:The serum levels of HMGB1 and IDO in ESCC group were significantly higher than those in control group (all P<0.01). Elevated serum HMGB1 and IDO levels were the independent risk factors for clinical progression of ESCC (all P<0.01), and their combined detection showed a higher predictive value for the clinical progression of ESCC (P<0.01). Serum HMGB1 and IDO levels were significantly correlated with T stage, N stage and clinical stage in ESCC patients (all P<0.05); Serum HMGB1 levels in ESCC group was negatively correlated with the absolute counts of CD3+ T cells, CD4+ T cells, B cells and NK cells in peripheral blood, and positively correlated with Treg cell percentage (all P<0.05); while serum IDO levels was negatively correlated with percentage and absolute count of CD3+ T cells, CD4+ T cells, and absolute counts of CD8+ T cells and B cells in peripheral blood,but positively correlated with the percentage of Treg cells (all P<0.05). Serum HMGB1 was positively correlated with IDO (P<0.01). The expression levels of IDO and p65 in KYSE30 and ECA109 cells and their culture supernatant in si-HMGB1 group were significantly lower than those in si-NC group and si-HMGB1+PMA group (all P<0.05). Conclusion: Serum HMGB1 and IDO are closely related to the clinical progression of ESCC and immune function of ESCC patients, and have the potential to serve as tumor markers and new immunotherapy targets of ESCC. HMGB1 may promote IDO expression through NF-κB signaling pathway, and dual-target combination therapy may achieve better efficacy.
9 Immunotherapy efficacy in 79 patients with malignant esophageal melanoma and the prognostic factors
2023, 30(7):612-615.
DOI: 10.3872/j.issn.1007-385X.2023.07.009
Abstract:
Objective: To explore clinical characteristics of patients with malignant esophageal melanoma (MEM) and analyze the efficacy and prognostic factors of PD-1 monoclonal antibody-based immunotherapy. Methods: Clinical data of patients with unresectable or metastatic MEM in the Department of Melanoma and Sarcoma of Peking University Cancer Hospital from May 2011 to June 2022 were collected, including basic information, pathological data, laboratory indicators, treatment patterns and survival situation. The clinical efficacy was evaluated using the Response Evaluation Criteria in Solid Tumors 1.1 (RECST 1.1). Kaplan-Meier curve was used for survival analysis, and univariate and multivariate COX regression analyses were used to screen prognostic factors. Results: A total of 79 MEM patients with complete data were enrolled, with a median age of 59.0. Most of the patients were accompanied by choking and dysphagia with the most common anatomic site of lower esophagus. There was a high mutant rate in NRAS and KIT, and the rate of LDH elevation accounted for 21.5%. Among them, 17 patients received chemotherapy, and 62 patients received PD-1 monoclonal antibody-based immunotherapy. The objective response rate (ORR) was 5.9% and 28.8%, and disease control rate ( DCR) was 35.3% and 72.9%, respectively. Overall survival (OS) in immune therapy group was significantly longer (13.2 months, 95%CI [9.5,16.9] months vs 7.0 months, 95%CI [0,16.7] months, P=0.019) than that in the chemotherapy group. Multivariate analysis showed that OS was significantly correlated with serum LDH, ECOG score, clinical symptoms and PD-1 monoclonal antibody-based immunotherapy (all P<0.05). Conclusion: MEM patients respond better to immune therapy, and elevated serum LDH, ECOG≥2, clinical symptoms at visiting might be the poor prognosis factors.
Objective: To explore clinical characteristics of patients with malignant esophageal melanoma (MEM) and analyze the efficacy and prognostic factors of PD-1 monoclonal antibody-based immunotherapy. Methods: Clinical data of patients with unresectable or metastatic MEM in the Department of Melanoma and Sarcoma of Peking University Cancer Hospital from May 2011 to June 2022 were collected, including basic information, pathological data, laboratory indicators, treatment patterns and survival situation. The clinical efficacy was evaluated using the Response Evaluation Criteria in Solid Tumors 1.1 (RECST 1.1). Kaplan-Meier curve was used for survival analysis, and univariate and multivariate COX regression analyses were used to screen prognostic factors. Results: A total of 79 MEM patients with complete data were enrolled, with a median age of 59.0. Most of the patients were accompanied by choking and dysphagia with the most common anatomic site of lower esophagus. There was a high mutant rate in NRAS and KIT, and the rate of LDH elevation accounted for 21.5%. Among them, 17 patients received chemotherapy, and 62 patients received PD-1 monoclonal antibody-based immunotherapy. The objective response rate (ORR) was 5.9% and 28.8%, and disease control rate ( DCR) was 35.3% and 72.9%, respectively. Overall survival (OS) in immune therapy group was significantly longer (13.2 months, 95%CI [9.5,16.9] months vs 7.0 months, 95%CI [0,16.7] months, P=0.019) than that in the chemotherapy group. Multivariate analysis showed that OS was significantly correlated with serum LDH, ECOG score, clinical symptoms and PD-1 monoclonal antibody-based immunotherapy (all P<0.05). Conclusion: MEM patients respond better to immune therapy, and elevated serum LDH, ECOG≥2, clinical symptoms at visiting might be the poor prognosis factors.
2023, 30(7):616-621.
DOI: 10.3872/j.issn.1007-385X.2023.07.010
Abstract:
酪蛋白激酶2（CK2）是一种高度保守的丝/苏氨酸蛋白激酶，它能够促进肿瘤细胞增殖、分化、侵袭和转移，并协助肿瘤细胞抗凋亡，亦有助于形成肿瘤免疫抑制微环境。近年来多项研究结果表明，CK2在乳腺癌细胞中呈过表达状态，是乳腺癌发生与发展过程中不可或缺的蛋白激酶。目前，CK2作为乳腺癌的新型治疗靶点，其在国内外的相关研究相对较少，但研发的各型CK2抑制剂已展现出抑制乳腺癌的巨大潜能。阐明CK2的结构和生物学功能及其在乳腺癌发展中的作用机制，以及CK2抑制剂的开发和临床应用现状，将有助于以CK2为靶点的乳腺癌靶向治疗。
酪蛋白激酶2（CK2）是一种高度保守的丝/苏氨酸蛋白激酶，它能够促进肿瘤细胞增殖、分化、侵袭和转移，并协助肿瘤细胞抗凋亡，亦有助于形成肿瘤免疫抑制微环境。近年来多项研究结果表明，CK2在乳腺癌细胞中呈过表达状态，是乳腺癌发生与发展过程中不可或缺的蛋白激酶。目前，CK2作为乳腺癌的新型治疗靶点，其在国内外的相关研究相对较少，但研发的各型CK2抑制剂已展现出抑制乳腺癌的巨大潜能。阐明CK2的结构和生物学功能及其在乳腺癌发展中的作用机制，以及CK2抑制剂的开发和临床应用现状，将有助于以CK2为靶点的乳腺癌靶向治疗。
2023, 30(7):622-627.
DOI: 10.3872/j.issn.1007-385X.2023.07.011
Abstract:
肿瘤引流淋巴结（TDLN）在肿瘤发生发展过程中起到介导肿瘤细胞转移和激发抗肿瘤免疫的双重作用。鉴于肿瘤免疫治疗的成功，需要对TDLN进行深入探讨以重新评估TDLN的手术切除与相关放疗。TDLN中有着与肿瘤微环境同样重要的特殊微环境，其中包括一系列免疫刺激和免疫抑制机制的对抗，并影响TDLN中包括T细胞、NK细胞在内的多种免疫细胞的活性及后续的抗肿瘤作用。TDLN间高度异质性也意味着需要对不同状态的TDLN采取系统化的研究以设计针对性的治疗措施。TDLN在肿瘤免疫中的重要地位使得可以通过靶向TDLN的方式达到提高疗效和减少毒副作用的效果。
肿瘤引流淋巴结（TDLN）在肿瘤发生发展过程中起到介导肿瘤细胞转移和激发抗肿瘤免疫的双重作用。鉴于肿瘤免疫治疗的成功，需要对TDLN进行深入探讨以重新评估TDLN的手术切除与相关放疗。TDLN中有着与肿瘤微环境同样重要的特殊微环境，其中包括一系列免疫刺激和免疫抑制机制的对抗，并影响TDLN中包括T细胞、NK细胞在内的多种免疫细胞的活性及后续的抗肿瘤作用。TDLN间高度异质性也意味着需要对不同状态的TDLN采取系统化的研究以设计针对性的治疗措施。TDLN在肿瘤免疫中的重要地位使得可以通过靶向TDLN的方式达到提高疗效和减少毒副作用的效果。
2023, 30(7):628-633.
DOI: 10.3872/j.issn.1007-385X.2023.07.012
Abstract:
巨噬细胞在实体瘤的发生发展中起着重要作用，基于巨噬细胞的实体瘤治疗也取得了阶段性的成果。与巨噬细胞相关的实体瘤治疗新策略主要包括：调节肿瘤相关巨噬细胞的数量和表型及构建嵌合抗原受体巨噬细胞（CAR-M），前者通过抑制巨噬细胞募集、耗竭肿瘤相关巨噬细胞（TAM）和重编程TAM等方式，减弱TAM对实体瘤的促进作用，增强TAM的抗肿瘤效应；后者基于CAR-T 的设计原理，使得巨噬细胞能够特异性识别肿瘤细胞表面抗原，并利用巨噬细胞较强的浸润能力，使得CAR-M能够进入实体瘤发挥抗肿瘤效应。目前，CAR-M疗法已在动物模型中获得了成功，相关临床研究正在进行，尚未获得明确的结论。TAM具有良好的组织浸润能力、吞噬活性和安全性，在实体瘤治疗中展现出广阔的前景，进一步研究TAM表型极化的机制、解决巨噬细胞来源问题、优化CAR-M的体外转染策略，将有效推动巨噬细胞在实体瘤治疗中的应用。
巨噬细胞在实体瘤的发生发展中起着重要作用，基于巨噬细胞的实体瘤治疗也取得了阶段性的成果。与巨噬细胞相关的实体瘤治疗新策略主要包括：调节肿瘤相关巨噬细胞的数量和表型及构建嵌合抗原受体巨噬细胞（CAR-M），前者通过抑制巨噬细胞募集、耗竭肿瘤相关巨噬细胞（TAM）和重编程TAM等方式，减弱TAM对实体瘤的促进作用，增强TAM的抗肿瘤效应；后者基于CAR-T 的设计原理，使得巨噬细胞能够特异性识别肿瘤细胞表面抗原，并利用巨噬细胞较强的浸润能力，使得CAR-M能够进入实体瘤发挥抗肿瘤效应。目前，CAR-M疗法已在动物模型中获得了成功，相关临床研究正在进行，尚未获得明确的结论。TAM具有良好的组织浸润能力、吞噬活性和安全性，在实体瘤治疗中展现出广阔的前景，进一步研究TAM表型极化的机制、解决巨噬细胞来源问题、优化CAR-M的体外转染策略，将有效推动巨噬细胞在实体瘤治疗中的应用。
2023, 30(7):634-638.
DOI: 10.3872/j.issn.1007-385X.2023.07.013
Abstract:
由于多发性骨髓瘤(MM)细胞及其微环境的高度异质性，导致其诊断和治疗方面面临巨大挑战。单细胞测序（sc-Seq）技术是一种以单个细胞为测试对象，分析其细胞内遗传信息的研究工具，近年来sc-Seq 在MM研究及治疗中广泛应用，为其更精准的诊断及个体化治疗作出了重要贡献。本文论述了近年来sc-Seq 技术在MM细胞异质性、免疫微环境、生物标志物及其在精准诊疗等方面应用的研究进展，为MM的基础与临床诊疗研究提供了参考依据。
由于多发性骨髓瘤(MM)细胞及其微环境的高度异质性，导致其诊断和治疗方面面临巨大挑战。单细胞测序（sc-Seq）技术是一种以单个细胞为测试对象，分析其细胞内遗传信息的研究工具，近年来sc-Seq 在MM研究及治疗中广泛应用，为其更精准的诊断及个体化治疗作出了重要贡献。本文论述了近年来sc-Seq 技术在MM细胞异质性、免疫微环境、生物标志物及其在精准诊疗等方面应用的研究进展，为MM的基础与临床诊疗研究提供了参考依据。
2023, 30(7):639-644.
DOI: 10.3872/j.issn.1007-385X.2023.07.014
Abstract:
近年来基因给药系统在肿瘤治疗中的研究备受关注。纳米载体具有增强药物靶向性、增加生物膜通透性、控制药物释放速度、提高生物利用度和可承载生物大分子等优点。由于肿瘤的快速增殖和代谢，形成了具有低pH、高水平谷胱甘肽、高水平活性氧、缺氧性、高表达酶和高水平ATP等特性的肿瘤微环境（TME）。基于肿瘤组织特异性微环境的基因给药纳米载体能同时提高基因药物的胞外稳定性、胞内释放能力和靶向性，可更大程度地提高药物的抗肿瘤作用、减少不良反应的发生。本文对基于TME的pH响应型、还原响应型、活性氧响应型、缺氧响应型、酶响应型、ATP响应型和多智能响应型纳米载体进行综述，总结各类型纳米载体的作用机制、制备、抗肿瘤效果及局限性。基因给药纳米载体可提高基因的转染效率，提高抗肿瘤作用，在抗肿瘤应用中有较好的前景。
近年来基因给药系统在肿瘤治疗中的研究备受关注。纳米载体具有增强药物靶向性、增加生物膜通透性、控制药物释放速度、提高生物利用度和可承载生物大分子等优点。由于肿瘤的快速增殖和代谢，形成了具有低pH、高水平谷胱甘肽、高水平活性氧、缺氧性、高表达酶和高水平ATP等特性的肿瘤微环境（TME）。基于肿瘤组织特异性微环境的基因给药纳米载体能同时提高基因药物的胞外稳定性、胞内释放能力和靶向性，可更大程度地提高药物的抗肿瘤作用、减少不良反应的发生。本文对基于TME的pH响应型、还原响应型、活性氧响应型、缺氧响应型、酶响应型、ATP响应型和多智能响应型纳米载体进行综述，总结各类型纳米载体的作用机制、制备、抗肿瘤效果及局限性。基因给药纳米载体可提高基因的转染效率，提高抗肿瘤作用，在抗肿瘤应用中有较好的前景。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.