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Volume 30,Issue 8,2023 Table of Contents
2023, 30(8):649-655.
DOI: 10.3872/j.issn.1007-385X.2023.08.001
Abstract:
Mammalian normal cells and tumor cells can release extracellular vesicles with 0.1-1 μm diameter when apoptosis or signal stimulation occurs. These signals stimulate the cytoskeleton changes to cause the plasma membrane to wrap the cellular contents and form extracellular vesicles, which are called tumor vesicles, which not only affect the biological characteristics of tumor cells but also have a profound impact on the tumor immune microenvironment. In addition to the biological effects, tumor vesicles can be used as a natural drug carrier, delivering therapeutic drugs to tumor cells and exerting anti-tumor effects. Follow-up studies have confirmed that drug-delivery tumor vesicles have a good anti-tumor activation effect in both innate and acquired immune responses. At present, drug-delivery tumor vesicles have entered the stage of clinical application. It has shown good application prospects in treating cholangiocarcinoma and malignant pleural effusion.
Mammalian normal cells and tumor cells can release extracellular vesicles with 0.1-1 μm diameter when apoptosis or signal stimulation occurs. These signals stimulate the cytoskeleton changes to cause the plasma membrane to wrap the cellular contents and form extracellular vesicles, which are called tumor vesicles, which not only affect the biological characteristics of tumor cells but also have a profound impact on the tumor immune microenvironment. In addition to the biological effects, tumor vesicles can be used as a natural drug carrier, delivering therapeutic drugs to tumor cells and exerting anti-tumor effects. Follow-up studies have confirmed that drug-delivery tumor vesicles have a good anti-tumor activation effect in both innate and acquired immune responses. At present, drug-delivery tumor vesicles have entered the stage of clinical application. It has shown good application prospects in treating cholangiocarcinoma and malignant pleural effusion.
ZHANG Tinglin
,
WU Lili
,
WANG Yu
,
ZHANG Zhuanzhuan
,
ZHOU Xuan
,
LI Meigui
,
YAN Zhenzhen
,
DING Xiuwen
,
LU Songwei
,
CHEN Cuimin
,
LIANG Hao
,
ZHANG Mengya
,
GAO Jie
2023, 30(8):656-664.
DOI: 10.3872/j.issn.1007-385X.2023.08.002
Abstract:
Objective: Crudlan (Cur) was loaded with manganese dioxide (MnO2) nanoparticle to construct a composite hydrogel MnO2@Cur (abbreviated as MGel) and its killing effect on melanoma B16-F10 cells was studied. Methods: Firstly, Cur hydrogel (Gel) was prepared by thermal induction method, and then MnO2 was physically loaded to construct MGel. The macroscopic and microscopic morphology of MGel was characterized and its mechanical properties, degradation properties and photothermal conversion properties were detected. The photothermal killing effect of MGel combined with PTT on mouse cutaneous melanoma B16-F10 cells was studied. Results: MGel is excellent in mechanical strength and degradation performance. Its tensile strength reached (127.97±3.60) kPa，and compressive strength reached (151.44±5.23) kPa; its degradation rate on the 28th day was about 58.17%. MGel obtained excellent photothermal conversion capability after loading MnO2 nanosheets with the average particle diameter of about 180 nm. After receiving 808 nm NIR illumination at 1.0 W/cm2 for 4 minutes, MGel loaded with 1.0 mg/mL MnO2 reached a maximum temperature of 50 ℃. Cytotoxicity experiments and Calcein-AM/PI fluorescence double staining showed that MGel combined with PTT killed B16-F10 melanoma cells effectively, and NIR illumination reduced the cell survival rate of the MGel group to (4.68±0.66)% (P<0.000 1). Conclusion: Composite hydrogel MGel has excellent mechanical strength, degradation performance and photothermal conversion capability. MGel combined with PTT can kill tumor cells effectively, which provides a novel strategy for the treatment of melanoma.
Objective: Crudlan (Cur) was loaded with manganese dioxide (MnO2) nanoparticle to construct a composite hydrogel MnO2@Cur (abbreviated as MGel) and its killing effect on melanoma B16-F10 cells was studied. Methods: Firstly, Cur hydrogel (Gel) was prepared by thermal induction method, and then MnO2 was physically loaded to construct MGel. The macroscopic and microscopic morphology of MGel was characterized and its mechanical properties, degradation properties and photothermal conversion properties were detected. The photothermal killing effect of MGel combined with PTT on mouse cutaneous melanoma B16-F10 cells was studied. Results: MGel is excellent in mechanical strength and degradation performance. Its tensile strength reached (127.97±3.60) kPa，and compressive strength reached (151.44±5.23) kPa; its degradation rate on the 28th day was about 58.17%. MGel obtained excellent photothermal conversion capability after loading MnO2 nanosheets with the average particle diameter of about 180 nm. After receiving 808 nm NIR illumination at 1.0 W/cm2 for 4 minutes, MGel loaded with 1.0 mg/mL MnO2 reached a maximum temperature of 50 ℃. Cytotoxicity experiments and Calcein-AM/PI fluorescence double staining showed that MGel combined with PTT killed B16-F10 melanoma cells effectively, and NIR illumination reduced the cell survival rate of the MGel group to (4.68±0.66)% (P<0.000 1). Conclusion: Composite hydrogel MGel has excellent mechanical strength, degradation performance and photothermal conversion capability. MGel combined with PTT can kill tumor cells effectively, which provides a novel strategy for the treatment of melanoma.
2023, 30(8):665-671.
DOI: 10.3872/j.issn.1007-385X.2023.08.003
Abstract:
Objective: To construct a recombinant immunotoxin targeting CD70 molecule, prepare PE38 and anti-CD70 nanobody recombinant proteins by expression and purification, and explore whether the recombinant proteins have killing activity on positive tumor cells with high expression of CD70 molecule through in vitro anti-tumor experiments. Methods: The CD70 nanobody Nb 2B3 gene fragment was ligated with the pET21a-PE38 gene fragment through a linker by genetic engineering, and the recombinant expression vector pET21a-Nb 2B3-PE38 was obtained and transformed into BL21 (DE3) competent cells for expression purification and identification. The binding activity of Nb 2B3-PE38 to CD70 molecule was detected by indirect ELISA and FACS. The in vitro killing activity of Nb 2B3-PE38 against renal clear cell carcinoma (786-O) with high expression of CD70 molecule was detected by MTT assay. The effect of Nb 2B3-PE38 on the apoptosis of 786-O cells was identified by Annexin Ⅴ-FITC/PI double staining.Results: The recombinant immunotoxin Nb 2B3-PE38 against CD70 nanobody was successfully constructed, and the recombinant protein with a purity higher than 90% was purified. The results of SDS-PAGE and WB showed that the target protein was correctly expressed and the molecular weight was 56 000. The purified Nb 2B3-PE38 could specifically bind to the recombinant CD70 antigen and the CD70 molecule on the surface of 786-O cells. 25 μg/mL Nb 2B3-PE38 has a highly significant killing effect on 786-O cells (P<0.001), and promotes apoptosis of 786-O cells (P<0.01). Its killing effect was stronger than that of positive control cisplatin (P<0.01). Conclusion: The immunotoxin Nb 2B3-PE38, which specifically targets CD70 molecules, was successfully prepared. This immunotoxin can effectively kill 786-O cells and induce apoptosis.
Objective: To construct a recombinant immunotoxin targeting CD70 molecule, prepare PE38 and anti-CD70 nanobody recombinant proteins by expression and purification, and explore whether the recombinant proteins have killing activity on positive tumor cells with high expression of CD70 molecule through in vitro anti-tumor experiments. Methods: The CD70 nanobody Nb 2B3 gene fragment was ligated with the pET21a-PE38 gene fragment through a linker by genetic engineering, and the recombinant expression vector pET21a-Nb 2B3-PE38 was obtained and transformed into BL21 (DE3) competent cells for expression purification and identification. The binding activity of Nb 2B3-PE38 to CD70 molecule was detected by indirect ELISA and FACS. The in vitro killing activity of Nb 2B3-PE38 against renal clear cell carcinoma (786-O) with high expression of CD70 molecule was detected by MTT assay. The effect of Nb 2B3-PE38 on the apoptosis of 786-O cells was identified by Annexin Ⅴ-FITC/PI double staining.Results: The recombinant immunotoxin Nb 2B3-PE38 against CD70 nanobody was successfully constructed, and the recombinant protein with a purity higher than 90% was purified. The results of SDS-PAGE and WB showed that the target protein was correctly expressed and the molecular weight was 56 000. The purified Nb 2B3-PE38 could specifically bind to the recombinant CD70 antigen and the CD70 molecule on the surface of 786-O cells. 25 μg/mL Nb 2B3-PE38 has a highly significant killing effect on 786-O cells (P<0.001), and promotes apoptosis of 786-O cells (P<0.01). Its killing effect was stronger than that of positive control cisplatin (P<0.01). Conclusion: The immunotoxin Nb 2B3-PE38, which specifically targets CD70 molecules, was successfully prepared. This immunotoxin can effectively kill 786-O cells and induce apoptosis.
HE Binli
,
LI Shanzhi
,
LI Yiquan
,
HAN Jicheng
,
LI Xiao
,
FANG Jinbo
,
JIN Ningyi
,
TIAN Tong
,
XUE Zhilong
,
ZHU Yilong
,
LI Zhuoxin
,
ZHU Guangze
2023, 30(8):672-680.
DOI: 10.3872/j.issn.1007-385X.2023.08.004
Abstract:
Objective: To study the effects of glabridin on malignant biological behaviors of lung adenocarcinoma A549 cells and its molecular mechanism. Methods: A549 cells and human normal lung epithelial BEAS-2B cells were cultured by conventional methods and then treated with different concentrations of glabridin and/or cisplatin. The effects of glabridin and cisplatin on the proliferative vitality of A549 and BEAS-2B cells were detected by crystal violet staining and CCK-8 method. Transwell cell assay and cell scratch assay were used to detect the effect of glabridin on the invasion and migration ability of A549 cells. Flow cytometry was conducted to detect the effect of glabridin on the apoptosis of A549 cells. 3D ultra-low adhesion plate culture method was applied to culture A549cells, and then the effects of glabridin on the proliferation of A549 cells and the expression of epithelial mesenchymal transition (EMT)-related proteins in A549 cells were detected using CCK-8 method and Western-blot, respectively. An A549 cell-transplanted tumor model was constructed to detect the effects of glabridin and cisplatin on the growth of transplanted tumors and the expression of EMT-related proteins in transplanted tumor tissues. Results: Glabridin and cisplatin significantly inhibited the proliferation (P<0.05 or P<0.01), cell migration (P<0.05 or P<0.01) and invasion (P<0.05 or P<0.01) of A549 cells in a dose-dependent manner. Glabridin could induce the apoptosis of A549 cells (P<0.01), inhibit the protein expression of N-cadherin, snail, and vimentin, but promote the protein expression of E-cadherin in A549 cells. Both glabridin and cisplatin could inhibit the growth of A549 cell-transplanted tumors,inhibit the protein expression of Ki67, N-cadherin, snail, and vimentin, and promote the protein expression of E-cadherin in transplanted tumor tissues. Conclusion: Glabridin can inhibit the proliferation, migration, and invasion of A549 cells, induce apoptosis of A549 cells, and inhibit EMT processes, thus exerting anti-tumor effect.
Objective: To study the effects of glabridin on malignant biological behaviors of lung adenocarcinoma A549 cells and its molecular mechanism. Methods: A549 cells and human normal lung epithelial BEAS-2B cells were cultured by conventional methods and then treated with different concentrations of glabridin and/or cisplatin. The effects of glabridin and cisplatin on the proliferative vitality of A549 and BEAS-2B cells were detected by crystal violet staining and CCK-8 method. Transwell cell assay and cell scratch assay were used to detect the effect of glabridin on the invasion and migration ability of A549 cells. Flow cytometry was conducted to detect the effect of glabridin on the apoptosis of A549 cells. 3D ultra-low adhesion plate culture method was applied to culture A549cells, and then the effects of glabridin on the proliferation of A549 cells and the expression of epithelial mesenchymal transition (EMT)-related proteins in A549 cells were detected using CCK-8 method and Western-blot, respectively. An A549 cell-transplanted tumor model was constructed to detect the effects of glabridin and cisplatin on the growth of transplanted tumors and the expression of EMT-related proteins in transplanted tumor tissues. Results: Glabridin and cisplatin significantly inhibited the proliferation (P<0.05 or P<0.01), cell migration (P<0.05 or P<0.01) and invasion (P<0.05 or P<0.01) of A549 cells in a dose-dependent manner. Glabridin could induce the apoptosis of A549 cells (P<0.01), inhibit the protein expression of N-cadherin, snail, and vimentin, but promote the protein expression of E-cadherin in A549 cells. Both glabridin and cisplatin could inhibit the growth of A549 cell-transplanted tumors,inhibit the protein expression of Ki67, N-cadherin, snail, and vimentin, and promote the protein expression of E-cadherin in transplanted tumor tissues. Conclusion: Glabridin can inhibit the proliferation, migration, and invasion of A549 cells, induce apoptosis of A549 cells, and inhibit EMT processes, thus exerting anti-tumor effect.
2023, 30(8):681-688.
DOI: 10.3872/j.issn.1007-385X.2023.08.005
Abstract:
Objective: To explore the possible mechanism of RAD18 affecting the proliferation of colorectal cancer cells and regulating the killing effect of NK cells on colorectal cells. Methods: Bioinformatics was used to analyze the expression of RAD18 andmiR-145-5p in colorectal cancer tissues and the regulatory relationship between them, and to analyze the RAD18 enrichment pathway.The expression of RAD18 and miR-145-5p in colorectal cancer cells was verified by qPCR. Dual-luciferase reporter gene assay verifiedthe regulatory relationship between miR-145-5p and RAD18. SW480 and HCT-15 cells were divided into si-RAD18 group and si-NC group according to the transfection; and SW480 cells were transfected inhibitor-NC+si-NC, miR-145-5p inhibitor+si-NC or miR-145-5pinhibitor+si-RAD18, respectively. The effects of knocking down miR-145-5p and/or RAD18 on cell proliferation and clonalization were detected by CCK-8 method and cloning formation experiments, respectively. The cells of each group were co-cultured with IL-2-activated NK92 cells, and the cytotoxicity, cytokine secretion, cell surface perforin and granzyme B expression of NK cells were detected by lactate dehydrogenase release assay, ELISA and immunofluorescence staining, respectively. Results: RAD18 was significantly over-expressed in colorectal cancer tissues and cells (all P<0.01). Silencing RAD18 can inhibit proliferation of colorectal cancer cells (P<0.05), promote NK cell viability, cytotoxicity, secretion of IFN- γ, TNF- α, GM-CSF and expression of perforin and granulozyme B (all P<0.05). In addition, dual-luciferase reporter assay verified the binding relationship between RAD18-3 'UTR and miR-145-5p, which is underexpressed in colorectal cancer tissues and cells (P<0.05 or P<0.01). miR-145-5p can down-regulate the expression of RAD18 by targeting (P<0.05), and over-expression of RAD18 can reverse the promotion effect of miR-145-5p over- expression on NK cell killing (all P<0.05). Conclusion: miR-145-5p can target down-regulation of RAD18 expression, and the miR-145-5p/RAD18 axis can affect the proliferation of colorectal cancer cells and the cytotoxic effect of NK cells.
Objective: To explore the possible mechanism of RAD18 affecting the proliferation of colorectal cancer cells and regulating the killing effect of NK cells on colorectal cells. Methods: Bioinformatics was used to analyze the expression of RAD18 andmiR-145-5p in colorectal cancer tissues and the regulatory relationship between them, and to analyze the RAD18 enrichment pathway.The expression of RAD18 and miR-145-5p in colorectal cancer cells was verified by qPCR. Dual-luciferase reporter gene assay verifiedthe regulatory relationship between miR-145-5p and RAD18. SW480 and HCT-15 cells were divided into si-RAD18 group and si-NC group according to the transfection; and SW480 cells were transfected inhibitor-NC+si-NC, miR-145-5p inhibitor+si-NC or miR-145-5pinhibitor+si-RAD18, respectively. The effects of knocking down miR-145-5p and/or RAD18 on cell proliferation and clonalization were detected by CCK-8 method and cloning formation experiments, respectively. The cells of each group were co-cultured with IL-2-activated NK92 cells, and the cytotoxicity, cytokine secretion, cell surface perforin and granzyme B expression of NK cells were detected by lactate dehydrogenase release assay, ELISA and immunofluorescence staining, respectively. Results: RAD18 was significantly over-expressed in colorectal cancer tissues and cells (all P<0.01). Silencing RAD18 can inhibit proliferation of colorectal cancer cells (P<0.05), promote NK cell viability, cytotoxicity, secretion of IFN- γ, TNF- α, GM-CSF and expression of perforin and granulozyme B (all P<0.05). In addition, dual-luciferase reporter assay verified the binding relationship between RAD18-3 'UTR and miR-145-5p, which is underexpressed in colorectal cancer tissues and cells (P<0.05 or P<0.01). miR-145-5p can down-regulate the expression of RAD18 by targeting (P<0.05), and over-expression of RAD18 can reverse the promotion effect of miR-145-5p over- expression on NK cell killing (all P<0.05). Conclusion: miR-145-5p can target down-regulation of RAD18 expression, and the miR-145-5p/RAD18 axis can affect the proliferation of colorectal cancer cells and the cytotoxic effect of NK cells.
2023, 30(8):689-694.
DOI: 10.3872/j.issn.1007-385X.2023.08.006
Abstract:
Objective: To investigate the mechanism of the inhibitory effects of dendrobium etract erianin (ER) on epithelial-mesenchymal transition (EMT) and angiogenesis of colorectal cancer HT29 cells based on the Hedgehog signal pathway. Methods: HT29 cells were separated into the blank control group, ER-L (25 μg/mL) group, ER-M (50 μg/mL) group, ER-H (75 μg/mL) group, and ER-H (75 μg/mL)+ PM (Hedgehog pathway activator, 1.5 μmol/L) group. MTT assay was applied to detect cell proliferative viability; the clonogenic ability of cells was detected by clonogenic assay; the abilities of cell migration and invasion were detected by scratch test and Transwell test; vascular mimicry test was applied to detect the ability of angiogenesis; WB assay was applied to detect the expressionof proteins related to EMT processes. Hedgehog signaling pathways, and mimic angiogenesis. Results: The proliferative viability of HT29 cells decreased gradually with the increase of ER concentration (P<0.05). Compared with the control group, cell clone formation rate, migration and invasion abilities,angiogenesis ability, the expressions of interstitial marker proteins (N-cadherin, vimentin), angiogenesis related proteins (VEGF, VE-cadherin) and hedgehog pathway related proteins (SHH, GLI1, SMO, c-Myc) in the various ER groups decreased, and the expressions of both epithelial marker protein (E-cadherin) and suppressor of fused protein (SUFU) in hedgehog pathway increased (all P<0.05). ER-H+PM group, to some extent, reversed the inhibition of ER on HT29 cell proliferation, EMT and angiogenesis(all P<0.05). Conclusion: ER can inhibit the proliferation, migration and invasion, EMT and angiogenesis of colorectal cancer HT29 cells, and its mechanism may be related to the inhibition of the activation of the Hedgehog signal pathway.
Objective: To investigate the mechanism of the inhibitory effects of dendrobium etract erianin (ER) on epithelial-mesenchymal transition (EMT) and angiogenesis of colorectal cancer HT29 cells based on the Hedgehog signal pathway. Methods: HT29 cells were separated into the blank control group, ER-L (25 μg/mL) group, ER-M (50 μg/mL) group, ER-H (75 μg/mL) group, and ER-H (75 μg/mL)+ PM (Hedgehog pathway activator, 1.5 μmol/L) group. MTT assay was applied to detect cell proliferative viability; the clonogenic ability of cells was detected by clonogenic assay; the abilities of cell migration and invasion were detected by scratch test and Transwell test; vascular mimicry test was applied to detect the ability of angiogenesis; WB assay was applied to detect the expressionof proteins related to EMT processes. Hedgehog signaling pathways, and mimic angiogenesis. Results: The proliferative viability of HT29 cells decreased gradually with the increase of ER concentration (P<0.05). Compared with the control group, cell clone formation rate, migration and invasion abilities,angiogenesis ability, the expressions of interstitial marker proteins (N-cadherin, vimentin), angiogenesis related proteins (VEGF, VE-cadherin) and hedgehog pathway related proteins (SHH, GLI1, SMO, c-Myc) in the various ER groups decreased, and the expressions of both epithelial marker protein (E-cadherin) and suppressor of fused protein (SUFU) in hedgehog pathway increased (all P<0.05). ER-H+PM group, to some extent, reversed the inhibition of ER on HT29 cell proliferation, EMT and angiogenesis(all P<0.05). Conclusion: ER can inhibit the proliferation, migration and invasion, EMT and angiogenesis of colorectal cancer HT29 cells, and its mechanism may be related to the inhibition of the activation of the Hedgehog signal pathway.
2023, 30(8):695-700.
DOI: 10.3872/j.issn.1007-385X.2023.08.007
Abstract:
Objective: To investigate the effects of carnosic acid (CA) on the proliferation, migration, and invasion of gastric cancer cells by regulating CXC chemokine receptor 7 (CXCR7)/CXC chemokine ligand 12 (CXCL12) axis. Methods: CCK-8 method was used to select appropriate concentrations of CA. Gastric cancer AGS cells were treated with CA at different concentrations (0, 5, 10, 20, 40, 80 μg/mL). AGS cells were divided into the control group (untreated AGS cells), CA group (20 μg/mL CA treatment), CA+ siCXCR7 group (siCXCR7 transfection+20 μg/mL CA treatment), CA+siNC group (siNC transfection+20 μg/mL CA treatment), CA+ vectorNC group (vectorNC transfection+20 μg/mL CA treatment), and CA+vectorCXCR7 group (vectorCXCR7 transfection+20 μg/mL CA treatment). The proliferation of AGS cells was detected by CCK-8 method; the expression levels of CXCR7 and CXCL12 mRNA in cells were detected by qPCR; and the invasion of cells was detected by Transwell assay; the change of cell migration ability was detected by scratch assay, and the expressions of cyclin D1, Bcl-2, CXCR7, CXCL12 and MMP-2 were detected by WB assay. Results: Different concentrations of CA were all able to inhibit the survival rate of AGS cells. When the concentration was 20 μg/mL, the survival rate of AGS cells was closed to 50%. 20 μg/mL CA was therefore chosen for subsequent research. Compared with the control group, the proliferation rate, invasion number, migration rate, cyclin D1, MMP-2, Bcl-2, CXCR7, CXCL12 mRNA and protein expression of the CA group decreased significantly (all P<0.05). Compared with the CA+siNC group, the proliferation rate, invasion number, migration rate, cyclin D1, MMP-2, Bcl-2, CXCR7, CXCL12 mRNA and protein expression in the CA+siCXCR7 group decreased significantly (all P<0.05). Compared with the CA+vectorNC group, the proliferation rate, invasion number, migration rate, cyclin D1, MMP-2, Bcl-2, CXCR7, CXCL12 mRNA and protein expression in the CA+vectorCXCR7 group increased significantly (all P<0.05). Conclusion: CA can inhibit the proliferation, migration, and invasion of AGS cells, which may be related to the inhibition of CXCR7/CXCL12 axis.
Objective: To investigate the effects of carnosic acid (CA) on the proliferation, migration, and invasion of gastric cancer cells by regulating CXC chemokine receptor 7 (CXCR7)/CXC chemokine ligand 12 (CXCL12) axis. Methods: CCK-8 method was used to select appropriate concentrations of CA. Gastric cancer AGS cells were treated with CA at different concentrations (0, 5, 10, 20, 40, 80 μg/mL). AGS cells were divided into the control group (untreated AGS cells), CA group (20 μg/mL CA treatment), CA+ siCXCR7 group (siCXCR7 transfection+20 μg/mL CA treatment), CA+siNC group (siNC transfection+20 μg/mL CA treatment), CA+ vectorNC group (vectorNC transfection+20 μg/mL CA treatment), and CA+vectorCXCR7 group (vectorCXCR7 transfection+20 μg/mL CA treatment). The proliferation of AGS cells was detected by CCK-8 method; the expression levels of CXCR7 and CXCL12 mRNA in cells were detected by qPCR; and the invasion of cells was detected by Transwell assay; the change of cell migration ability was detected by scratch assay, and the expressions of cyclin D1, Bcl-2, CXCR7, CXCL12 and MMP-2 were detected by WB assay. Results: Different concentrations of CA were all able to inhibit the survival rate of AGS cells. When the concentration was 20 μg/mL, the survival rate of AGS cells was closed to 50%. 20 μg/mL CA was therefore chosen for subsequent research. Compared with the control group, the proliferation rate, invasion number, migration rate, cyclin D1, MMP-2, Bcl-2, CXCR7, CXCL12 mRNA and protein expression of the CA group decreased significantly (all P<0.05). Compared with the CA+siNC group, the proliferation rate, invasion number, migration rate, cyclin D1, MMP-2, Bcl-2, CXCR7, CXCL12 mRNA and protein expression in the CA+siCXCR7 group decreased significantly (all P<0.05). Compared with the CA+vectorNC group, the proliferation rate, invasion number, migration rate, cyclin D1, MMP-2, Bcl-2, CXCR7, CXCL12 mRNA and protein expression in the CA+vectorCXCR7 group increased significantly (all P<0.05). Conclusion: CA can inhibit the proliferation, migration, and invasion of AGS cells, which may be related to the inhibition of CXCR7/CXCL12 axis.
2023, 30(8):701-706.
DOI: 10.3872/j.issn.1007-385X.2023.08.008
Abstract:
Objective: To investigate the killing effects of Epstein-Barr virus nuclear antigen 1 (EBNA1) mRNA-modified DC (EBNA1-DC) -induced lymphocytes combined with methylation inhibitor 5-Aza-CdR on nasopharyngeal carcinoma C666-1 cells.Methods: EBNA1-pCDNA3.1 plasmid was used as a template, and EBNA1 mRNA was obtained by in vitro transcription.Subsequently, EBNA1 mRNA was transfected into dendritic cells derived from peripheral blood from healthy donors by liposome toconstruct EBNA1-DC vaccine. Flow cytometry was used to detect the transfected DC phenotype and the apoptosis of C666-1 cells after 5-Aza-CdR treatment. Real-time cell analysis was used to detect the specific antitumor activity of EBNA1-DC vaccine-induced lymphocytes combined with 5-Aza-CdR. Results: The positive rate of EBNA1on the surface of EBNA1-DC after transfection with EBNA1 mRNA was (59.3±5.85) %. The expression of HLA-DR was significantly higher than that of untransfected DC ([84.9±5.5]% vs [68.0±5.8]% , P=0.026). The expression of CD80 was significantly improved from (88.2±3.9)% to (61.1±4.4)% (P=0.015). The apoptosis of C666-1 cells treated with low-dose 5-Aza-CdR was not significantly different from that of untreated cells. The expression of IRF7 gene in C666-1 cells pretreated with low-dos 5-Aza-CdR was significantly higher than that in untreated cells (P=0.000 1).Lymphocytes induced by EBNA1-DC had stronger specific killing activity against EBV+C666-1 cells compared with untransfected DC (P=0.049). C666-1 cells pretreated with low dose 5-Aza-CdR were more sensitive to specific immune killing induced by EBNA1-DC (P=0.019). Conclusion: The combination of 5-Aza-CdR and EBNA1-DC vaccine can significantly enhance the specific immune killing effect on C666-1 cells. This study provides the preliminary research basis for the development of the mRNA-DC vaccine and its application in clinical comprehensive therapy.
Objective: To investigate the killing effects of Epstein-Barr virus nuclear antigen 1 (EBNA1) mRNA-modified DC (EBNA1-DC) -induced lymphocytes combined with methylation inhibitor 5-Aza-CdR on nasopharyngeal carcinoma C666-1 cells.Methods: EBNA1-pCDNA3.1 plasmid was used as a template, and EBNA1 mRNA was obtained by in vitro transcription.Subsequently, EBNA1 mRNA was transfected into dendritic cells derived from peripheral blood from healthy donors by liposome toconstruct EBNA1-DC vaccine. Flow cytometry was used to detect the transfected DC phenotype and the apoptosis of C666-1 cells after 5-Aza-CdR treatment. Real-time cell analysis was used to detect the specific antitumor activity of EBNA1-DC vaccine-induced lymphocytes combined with 5-Aza-CdR. Results: The positive rate of EBNA1on the surface of EBNA1-DC after transfection with EBNA1 mRNA was (59.3±5.85) %. The expression of HLA-DR was significantly higher than that of untransfected DC ([84.9±5.5]% vs [68.0±5.8]% , P=0.026). The expression of CD80 was significantly improved from (88.2±3.9)% to (61.1±4.4)% (P=0.015). The apoptosis of C666-1 cells treated with low-dose 5-Aza-CdR was not significantly different from that of untreated cells. The expression of IRF7 gene in C666-1 cells pretreated with low-dos 5-Aza-CdR was significantly higher than that in untreated cells (P=0.000 1).Lymphocytes induced by EBNA1-DC had stronger specific killing activity against EBV+C666-1 cells compared with untransfected DC (P=0.049). C666-1 cells pretreated with low dose 5-Aza-CdR were more sensitive to specific immune killing induced by EBNA1-DC (P=0.019). Conclusion: The combination of 5-Aza-CdR and EBNA1-DC vaccine can significantly enhance the specific immune killing effect on C666-1 cells. This study provides the preliminary research basis for the development of the mRNA-DC vaccine and its application in clinical comprehensive therapy.
2023, 30(8):707-714.
DOI: 10.3872/j.issn.1007-385X.2023.08.009
Abstract:
Objective: To study the influence of circRNA nei-like DNA glucoamylase 3 (circNEIL3) and microRNA (miR)-4784 on the proliferation, migration, and invasion of breast cancer MDA-MB-231 cells. Methods: The cancer tissues and paired paracancerous tissues surgically removed from 45 breast cancer patients histopathologically diagnosed with breast cancer in Jinan Hospital of Integrated Traditional Chinese and Western Medicine from January 2018 to December 2019 were collected. qPCR were applied to measure the relative levels of circNEIL3 and miR-4784 in breast cancer tissues and paracancerous tissues. circNEIL3 small interfering RNA (si-circNEIL3), miR-4784 mimics, and si-circNEIL3+miR-4784 inhibitors were transfected into MDA-MB-231 breast cancer cells, respectively. CCK-8 assay, plate cloning assay, scratch healing assay, and Transwell assay were performed to detect the effects of circNEIL3 and miR-4784 expressions on the cell viability, clone formation, migration and invasion. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and RNA pull-down assay were used to detect the interactions between circNEIL3 and miR-4784. Results: The relative level of circNEIL3 in breast cancer tissues was significantly higher than that in adjacent tissues (P<0.05) while the relative level of miR-4784 was significantly lower than that in adjacent tissues (P<0.05). Interference with circNEIL3 significantly reduced the viability, clonal formation numbers, wound healing rate, and invasion numbers of MDA-MB-231 cells (P< 0.05). Overexpression of miR-4784 significantly reduced the viability, clonal formation numbers, wound healing rate, and invasion numbers of MDA-MB-231 cells (P<0.05). Dual luciferase reporter gene assay, RIP, and RNA pull-down assay confirmed that circNEIL3 directly binds to miR-4784. Interference with circNEIL3 significantly up-regulated miR-4784 expression (P<0.05), and overexpression of circNEIL3 significantly down-regulated miR-4784 expression (P<0.05). miR-4784 inhibition partially reversed the inhibitory effect of interference with circNEIL3 on the viability, colony formation, migration, and invasion of MDA-MB-231 cells (P<0.05). Conclusion: Interference with circNEIL3 inhibits the proliferation, migration, and invasion of breast cancer cells by targeting and up-regulating miR-4784 expression.
Objective: To study the influence of circRNA nei-like DNA glucoamylase 3 (circNEIL3) and microRNA (miR)-4784 on the proliferation, migration, and invasion of breast cancer MDA-MB-231 cells. Methods: The cancer tissues and paired paracancerous tissues surgically removed from 45 breast cancer patients histopathologically diagnosed with breast cancer in Jinan Hospital of Integrated Traditional Chinese and Western Medicine from January 2018 to December 2019 were collected. qPCR were applied to measure the relative levels of circNEIL3 and miR-4784 in breast cancer tissues and paracancerous tissues. circNEIL3 small interfering RNA (si-circNEIL3), miR-4784 mimics, and si-circNEIL3+miR-4784 inhibitors were transfected into MDA-MB-231 breast cancer cells, respectively. CCK-8 assay, plate cloning assay, scratch healing assay, and Transwell assay were performed to detect the effects of circNEIL3 and miR-4784 expressions on the cell viability, clone formation, migration and invasion. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and RNA pull-down assay were used to detect the interactions between circNEIL3 and miR-4784. Results: The relative level of circNEIL3 in breast cancer tissues was significantly higher than that in adjacent tissues (P<0.05) while the relative level of miR-4784 was significantly lower than that in adjacent tissues (P<0.05). Interference with circNEIL3 significantly reduced the viability, clonal formation numbers, wound healing rate, and invasion numbers of MDA-MB-231 cells (P< 0.05). Overexpression of miR-4784 significantly reduced the viability, clonal formation numbers, wound healing rate, and invasion numbers of MDA-MB-231 cells (P<0.05). Dual luciferase reporter gene assay, RIP, and RNA pull-down assay confirmed that circNEIL3 directly binds to miR-4784. Interference with circNEIL3 significantly up-regulated miR-4784 expression (P<0.05), and overexpression of circNEIL3 significantly down-regulated miR-4784 expression (P<0.05). miR-4784 inhibition partially reversed the inhibitory effect of interference with circNEIL3 on the viability, colony formation, migration, and invasion of MDA-MB-231 cells (P<0.05). Conclusion: Interference with circNEIL3 inhibits the proliferation, migration, and invasion of breast cancer cells by targeting and up-regulating miR-4784 expression.
2023, 30(8):715-719.
DOI: 10.3872/j.issn.1007-385X.2023.08.010
Abstract:
晚期不可切除肝细胞癌（HCC）是全球病死率最高的恶性肿瘤之一，其治疗方式有限。近年来，免疫检查点抑制剂治疗已成为HCC的主要治疗手段，显著延长了患者的生存期，但由于HCC特殊的免疫微环境，大部分患者仍存在治疗抵抗。肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）在HCC微环境的“再教育”下，浸润数量最多、亚型独特、功能复杂，不仅参与肿瘤的发生与发展，也能通过改变自身分子表达、调控其他细胞功能、重塑肿瘤微环境、搭建免疫耐受屏障等多种方式，以“核心调控者”的身份促使免疫治疗抵抗。因此，削减TAM的募集、提高抗瘤样TAM的比例或以其为基点协同其他治疗方式改善免疫抑制性微环境等针对性策略，能极大地提高HCC的免疫治疗效果。
晚期不可切除肝细胞癌（HCC）是全球病死率最高的恶性肿瘤之一，其治疗方式有限。近年来，免疫检查点抑制剂治疗已成为HCC的主要治疗手段，显著延长了患者的生存期，但由于HCC特殊的免疫微环境，大部分患者仍存在治疗抵抗。肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）在HCC微环境的“再教育”下，浸润数量最多、亚型独特、功能复杂，不仅参与肿瘤的发生与发展，也能通过改变自身分子表达、调控其他细胞功能、重塑肿瘤微环境、搭建免疫耐受屏障等多种方式，以“核心调控者”的身份促使免疫治疗抵抗。因此，削减TAM的募集、提高抗瘤样TAM的比例或以其为基点协同其他治疗方式改善免疫抑制性微环境等针对性策略，能极大地提高HCC的免疫治疗效果。
2023, 30(8):720-725.
DOI: 10.3872/j.issn.1007-385X.2023.08.011
Abstract:
多发性骨髓瘤（MM）是一种浆细胞的恶性增殖性疾病。在MM的发生发展过程中，骨髓微环境（BMME）的作用不容忽视。通过产生免疫抑制细胞、效应细胞功能失调和产生细胞因子和代谢物等抑制抗肿瘤免疫等方式，诱发免疫微环境逐渐失衡，最终导致肿瘤的发生与发展。因此，深入研究BMME可增强人们对MM的认识，对于MM的治疗亦有重要意义。基于近年来BMME的研究进展，笔者论述了抗MM免疫细胞和促MM免疫细胞的作用及其机制，探讨了MM免疫微环境对免疫治疗效果的影响，并展望了未来MM的免疫治疗方法，为探索新的MM的免疫治疗手段提供了方向。
多发性骨髓瘤（MM）是一种浆细胞的恶性增殖性疾病。在MM的发生发展过程中，骨髓微环境（BMME）的作用不容忽视。通过产生免疫抑制细胞、效应细胞功能失调和产生细胞因子和代谢物等抑制抗肿瘤免疫等方式，诱发免疫微环境逐渐失衡，最终导致肿瘤的发生与发展。因此，深入研究BMME可增强人们对MM的认识，对于MM的治疗亦有重要意义。基于近年来BMME的研究进展，笔者论述了抗MM免疫细胞和促MM免疫细胞的作用及其机制，探讨了MM免疫微环境对免疫治疗效果的影响，并展望了未来MM的免疫治疗方法，为探索新的MM的免疫治疗手段提供了方向。
2023, 30(8):726-732.
DOI: 10.3872/j.issn.1007-385X.2023.08.012
Abstract:
CD99是一种主要定位在细胞膜上的Ⅰ型跨膜糖蛋白，参与调控正常组织细胞的分化、黏附、迁移和蛋白运输等功能。CD99蛋白在大多数正常组织中表达较低，但在一些恶性肿瘤，尤其是尤因肉瘤（EWS）中高表达，可作为EWS的辅助诊断和预后标志物。高表达的CD99蛋白可通过多种机制促进EWS的发生发展，例如与配体结合、影响外泌体相关的miRNA分泌和调控下游信号通路等。作为在EWS细胞中高特异性表达并发挥促癌作用的膜蛋白，CD99 是理想的肿瘤治疗靶点，已有研究尝试开发通过靶向CD99治疗EWS的方法，包括CD99中和抗体、小分子抑制剂和肿瘤疫苗等。尽管目前靶向CD99治疗EWS还处于临床前研究阶段，但已展现出很好的治疗前景。因此，针对CD99的靶点干预可能为EWS治疗提供新策略。
CD99是一种主要定位在细胞膜上的Ⅰ型跨膜糖蛋白，参与调控正常组织细胞的分化、黏附、迁移和蛋白运输等功能。CD99蛋白在大多数正常组织中表达较低，但在一些恶性肿瘤，尤其是尤因肉瘤（EWS）中高表达，可作为EWS的辅助诊断和预后标志物。高表达的CD99蛋白可通过多种机制促进EWS的发生发展，例如与配体结合、影响外泌体相关的miRNA分泌和调控下游信号通路等。作为在EWS细胞中高特异性表达并发挥促癌作用的膜蛋白，CD99 是理想的肿瘤治疗靶点，已有研究尝试开发通过靶向CD99治疗EWS的方法，包括CD99中和抗体、小分子抑制剂和肿瘤疫苗等。尽管目前靶向CD99治疗EWS还处于临床前研究阶段，但已展现出很好的治疗前景。因此，针对CD99的靶点干预可能为EWS治疗提供新策略。
2023, 30(8):733-738.
DOI: 10.3872/j.issn.1007-385X.2023.08.013
Abstract:
肿瘤免疫微环境（TIME）及其对肺腺癌患者预后和治疗的影响已成为近年来肺腺癌研究的新热点。为了明确TIME的动态变化和其中各个组分对治疗和预后的影响，本综述分析近年来利用单细胞RNA测序（scRNA-seq）技术在肺腺癌中进行的TIME相关研究，发现肿瘤细胞、免疫细胞、各种调控因子及其在TIME中的相互作用影响着肺癌的发生、发展和治疗结果。TIME的异质性受各种癌细胞、免疫细胞、细胞因子/趋化因子环境、细胞毒活性或免疫抑制因子等影响。与TIME异质性相关的细胞亚型的特定组成可能促进或阻碍机体对免疫治疗的反应，并影响患者预后。目前，已发现多种标志物，包括差异表达基因、特异性细胞标志物、细胞性质相关标志物，均可对患者预后分层或预测治疗效果。另外，多种亚型的免疫细胞亦可对肺腺癌免疫治疗耐药性进行分层和预测。本文综述了TIME对预后分层、疗效预测和耐药性分层的相关标志物或模型，希望对未来肺腺癌治疗策略的制定和生物标志物的开发提供启发。
肿瘤免疫微环境（TIME）及其对肺腺癌患者预后和治疗的影响已成为近年来肺腺癌研究的新热点。为了明确TIME的动态变化和其中各个组分对治疗和预后的影响，本综述分析近年来利用单细胞RNA测序（scRNA-seq）技术在肺腺癌中进行的TIME相关研究，发现肿瘤细胞、免疫细胞、各种调控因子及其在TIME中的相互作用影响着肺癌的发生、发展和治疗结果。TIME的异质性受各种癌细胞、免疫细胞、细胞因子/趋化因子环境、细胞毒活性或免疫抑制因子等影响。与TIME异质性相关的细胞亚型的特定组成可能促进或阻碍机体对免疫治疗的反应，并影响患者预后。目前，已发现多种标志物，包括差异表达基因、特异性细胞标志物、细胞性质相关标志物，均可对患者预后分层或预测治疗效果。另外，多种亚型的免疫细胞亦可对肺腺癌免疫治疗耐药性进行分层和预测。本文综述了TIME对预后分层、疗效预测和耐药性分层的相关标志物或模型，希望对未来肺腺癌治疗策略的制定和生物标志物的开发提供启发。
2023, 30(8):739-741.
DOI: 10.3872/j.issn.1007-385X.2023.08.014
Abstract:
晚期结直肠癌预后差，总生存期短，其靶向药物的选择依赖于基因检测的结果。部分罕见的基因突变如间变性淋巴瘤激酶（anaplastic lymphoma kinase，ALK）融合基因突变在肠癌中发生率仅为0.4%~1%，其药物治疗的模式仍存在争议。本文报告1例晚期右半结肠癌合并ALK融合基因突变的患者，该患者在贝伐珠单抗靶向联合静脉化疗的标准治疗基础上同时接受ALK抑制剂阿来替尼治疗，第3周期化疗时患者临床症状明显缓解，肿瘤指标较前下降，胸部CT提示肺部病灶较前改善，取得了显著的临床获益。尽管ALK融合基因突变在结直肠癌中罕见，但其治疗模式的选择仍需要临床医生探索。
晚期结直肠癌预后差，总生存期短，其靶向药物的选择依赖于基因检测的结果。部分罕见的基因突变如间变性淋巴瘤激酶（anaplastic lymphoma kinase，ALK）融合基因突变在肠癌中发生率仅为0.4%~1%，其药物治疗的模式仍存在争议。本文报告1例晚期右半结肠癌合并ALK融合基因突变的患者，该患者在贝伐珠单抗靶向联合静脉化疗的标准治疗基础上同时接受ALK抑制剂阿来替尼治疗，第3周期化疗时患者临床症状明显缓解，肿瘤指标较前下降，胸部CT提示肺部病灶较前改善，取得了显著的临床获益。尽管ALK融合基因突变在结直肠癌中罕见，但其治疗模式的选择仍需要临床医生探索。
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.