Mobile website
Volume 31,Issue 2,2024 Table of Contents
2024, 31(2):113-120.
DOI: 10.3872/j.issn.1007-385X.2024.02.001
Abstract:
Leukemia inhibitory factor （LIF） is a pleiotropic cytokine of the interleukin-6 （IL-6） family, which was first identified as being able to inhibit the proliferation of mouse myeloid M1 leukemia cells and induce their terminal differentiation. LIF is widely involved in the regeneration of organs, neural development, and immune regulation, and also plays an important role in the development of tumors. In contrast to its inhibitory effect on the proliferation of leukemia cells, LIF typically facilitates the progression of many solid tumors. Elevated expression of LIF has been observed to contribute to the development, metastasis, treatment resistance, and evasion of immune response in tumors, hence correlating with a poor prognosis in patients. Focusing on the functions of LIF physiology and pathology and the holistic nature of the signaling pathways it regulates and searching for new targeted drugs is important for the development of targeted therapeutic strategies for the LIF pathway.
Leukemia inhibitory factor （LIF） is a pleiotropic cytokine of the interleukin-6 （IL-6） family, which was first identified as being able to inhibit the proliferation of mouse myeloid M1 leukemia cells and induce their terminal differentiation. LIF is widely involved in the regeneration of organs, neural development, and immune regulation, and also plays an important role in the development of tumors. In contrast to its inhibitory effect on the proliferation of leukemia cells, LIF typically facilitates the progression of many solid tumors. Elevated expression of LIF has been observed to contribute to the development, metastasis, treatment resistance, and evasion of immune response in tumors, hence correlating with a poor prognosis in patients. Focusing on the functions of LIF physiology and pathology and the holistic nature of the signaling pathways it regulates and searching for new targeted drugs is important for the development of targeted therapeutic strategies for the LIF pathway.
2024, 31(2):121-127.
DOI: 10.3872/j.issn.1007-385X.2024.02.002
Abstract:
Tertiary lymphoid structure (TLS) refers to ectopic lymphoid tissue formed at long-term inflammatory sites in non-physiological conditions. It is a lymphocyte aggregate constructed on the fibroblast network and contains two important structural regions-the T cell region and the follicular B cell region. TLS exists in different mature states in tumors, and finally forms germinal centers. Germinal centers contain T follicular helper cells and follicular dendritic cells, and are closely related to B cells. Recent studies have identified the main features, biomarkers and detection methods of newborn TLS, further expounding the mechanism that TLSs can enhance the effect of anti-tumor immune response by stimulating related tumor antigens and regulating lymphocyte infiltration. Studies on the relevance between TLSs and clinical benefits of cancer patients show that TLSs can be used as a good prognosis and predictive factor for biotherapies including immunotherapy. Several methods to induce TLS formation are currently being developed, including the use of chemokines, cytokines, antibodies, antigen presenting cells or synthetic scaffolds. The regimen of inducing TLS neogenesis in cold tumors and hot tumors in combination with therapeutic agents for inflammatory environment suppression or immune checkpoint inhibitors, represents a new hope for cancer treatment.
Tertiary lymphoid structure (TLS) refers to ectopic lymphoid tissue formed at long-term inflammatory sites in non-physiological conditions. It is a lymphocyte aggregate constructed on the fibroblast network and contains two important structural regions-the T cell region and the follicular B cell region. TLS exists in different mature states in tumors, and finally forms germinal centers. Germinal centers contain T follicular helper cells and follicular dendritic cells, and are closely related to B cells. Recent studies have identified the main features, biomarkers and detection methods of newborn TLS, further expounding the mechanism that TLSs can enhance the effect of anti-tumor immune response by stimulating related tumor antigens and regulating lymphocyte infiltration. Studies on the relevance between TLSs and clinical benefits of cancer patients show that TLSs can be used as a good prognosis and predictive factor for biotherapies including immunotherapy. Several methods to induce TLS formation are currently being developed, including the use of chemokines, cytokines, antibodies, antigen presenting cells or synthetic scaffolds. The regimen of inducing TLS neogenesis in cold tumors and hot tumors in combination with therapeutic agents for inflammatory environment suppression or immune checkpoint inhibitors, represents a new hope for cancer treatment.
YANG Jing
,
ZHANG Weiguang
,
LI Chencheng
,
LIU Xixi
,
HU Zhongxiao
,
WANG Fengnan
,
Chen Biqing
,
TIAN Fang
,
ZHANG Xiaoli
,
JIANATI Reaila
,
ZHU Xuejun
2024, 31(2):128-134.
DOI: 10.3872/j.issn.1007-385X.2024.02.003
Abstract:
Objective: To establish an animal model of cytokine release syndrome (CRS) by intraperitoneal injection of mouse IFN-γ adenovirus (Ad-mIFN- γ) into C57Bl/6J mice. Methods: Ad-mIFN- γ and Ad-LacZ control adenoviral vectors were constructed, and mouse peritoneal macrophages were transfected with MOI=100 in vitro. The effect on mIFN-γ secretion levels of cells were detected by flow cytometry. Forty female C57Bl/6J mice were divided into the control group, the vector control group, and the low-, medium-, and high-dose virus groups (8 mice in each group) according to the random number table method. The mice were intraperitoneally injected with PBS (200 μL/piece), Ad-lacZ (2×107 PFU/piece), Ad-mIFN-γ (5×106 PFU/piece), Ad-mIFN-γ (1.5×107 PFU/piece) and Ad-mIFN-γ (2×107 PFU/piece), respectively. The body weight and survival of the mice were observed daily. On the third day, flow cytometry was used to detect the proportion of monocytes (CD11b+) and macrophages (CD11b+/CD86+) in the peripheral blood and the spleen, and the proportion of CD11b+ monocytes in the spleen was detected by immunofluorescence staining. On the 9th day, flow cytometry was used to detect the secretion of cytokines in the serum of the mice. On the 14th day, the mice were sacrificed by cervical dislocation, and H-E staining was used to observe the pathological and histological changes of the liver, spleen, lungs and kidneys of the mice. Results: Ad-mIFN-γ infected mouse peritoneal macrophages in vitro, and the level of mIFN-γ secreted by macrophages was detected to reach a peak of (118.34±2.90) pg/mL on the third day, and the secretion level of mIFN-γ continued to be high for one week; while the Ad-lacZ control group secreted a lower level of IFN- γ, with a value of (0.17±0.08) pg/mL on the third day. After intraperitoneal injection of Ad-mIFN-γ, no mice died in the low- and medium-dose virus groups within 14 days, and the body weight of the mice in the high-dose virus group continued to decrease (P<0.001). On the third day, the proportions of monocytes and macrophages in the peripheral blood and spleen tissues of the mice in the high-dose virus group were significantly higher than those in the control group and the medium- dose group (P<0.05 or P<0.01). On the 9th day, the serum levels of mIFN-γ, IL-6, monocyte chemotactic protein-1 (MCP-1), IL-1 and TNF-α in the low-, medium- and high-dose groups increased significantly (P<0.001). Within 10 days, the mortality rate of the mice in the high-dose virus group reached 100%. Histopathological examination showed significant damage in the liver, spleen, lung and kidney tissues of the mice in the high-dose virus group. Conclusion: Mouse primary peritoneal macrophages can rapidly secrete mIFN-γ after infection with Ad-mIFN-γ in vitro. Intraperitoneal injection of high-dose Ad-mIFN-γ (2×107 PFU/piece) resulted in typical CRS manifestations in the mice, which can be used as an animal model for CAR-T cell therapy-induced CRS.
Objective: To establish an animal model of cytokine release syndrome (CRS) by intraperitoneal injection of mouse IFN-γ adenovirus (Ad-mIFN- γ) into C57Bl/6J mice. Methods: Ad-mIFN- γ and Ad-LacZ control adenoviral vectors were constructed, and mouse peritoneal macrophages were transfected with MOI=100 in vitro. The effect on mIFN-γ secretion levels of cells were detected by flow cytometry. Forty female C57Bl/6J mice were divided into the control group, the vector control group, and the low-, medium-, and high-dose virus groups (8 mice in each group) according to the random number table method. The mice were intraperitoneally injected with PBS (200 μL/piece), Ad-lacZ (2×107 PFU/piece), Ad-mIFN-γ (5×106 PFU/piece), Ad-mIFN-γ (1.5×107 PFU/piece) and Ad-mIFN-γ (2×107 PFU/piece), respectively. The body weight and survival of the mice were observed daily. On the third day, flow cytometry was used to detect the proportion of monocytes (CD11b+) and macrophages (CD11b+/CD86+) in the peripheral blood and the spleen, and the proportion of CD11b+ monocytes in the spleen was detected by immunofluorescence staining. On the 9th day, flow cytometry was used to detect the secretion of cytokines in the serum of the mice. On the 14th day, the mice were sacrificed by cervical dislocation, and H-E staining was used to observe the pathological and histological changes of the liver, spleen, lungs and kidneys of the mice. Results: Ad-mIFN-γ infected mouse peritoneal macrophages in vitro, and the level of mIFN-γ secreted by macrophages was detected to reach a peak of (118.34±2.90) pg/mL on the third day, and the secretion level of mIFN-γ continued to be high for one week; while the Ad-lacZ control group secreted a lower level of IFN- γ, with a value of (0.17±0.08) pg/mL on the third day. After intraperitoneal injection of Ad-mIFN-γ, no mice died in the low- and medium-dose virus groups within 14 days, and the body weight of the mice in the high-dose virus group continued to decrease (P<0.001). On the third day, the proportions of monocytes and macrophages in the peripheral blood and spleen tissues of the mice in the high-dose virus group were significantly higher than those in the control group and the medium- dose group (P<0.05 or P<0.01). On the 9th day, the serum levels of mIFN-γ, IL-6, monocyte chemotactic protein-1 (MCP-1), IL-1 and TNF-α in the low-, medium- and high-dose groups increased significantly (P<0.001). Within 10 days, the mortality rate of the mice in the high-dose virus group reached 100%. Histopathological examination showed significant damage in the liver, spleen, lung and kidney tissues of the mice in the high-dose virus group. Conclusion: Mouse primary peritoneal macrophages can rapidly secrete mIFN-γ after infection with Ad-mIFN-γ in vitro. Intraperitoneal injection of high-dose Ad-mIFN-γ (2×107 PFU/piece) resulted in typical CRS manifestations in the mice, which can be used as an animal model for CAR-T cell therapy-induced CRS.
2024, 31(2):135-145.
DOI: 10.3872/j.issn.1007-385X.2024.02.004
Abstract:
Objective: To screen the upstream miRNAs and lncRNAs of EZH2 gene, analyze their expressions in gastric cancer cells,verify the targeting relationship between them, and discuss their effects on the proliferation, migration and apoptosis of gastric cancer cells. Methods: The upstream miRNA (has-miR-450b-5p) of EZH2 was queried, analyzed and screened by ENCORI, miRDB and Target Scan databases, and the upstream lncRNA (lncRNA NEAT1) of hsa-miR-450b-5p was screened by ENCORI database and DAINA database. The binding sites between hsa-miR-450b-5p, lncRNA NEAT1 and EZH2 were predicted. Dual-luciferase reporter assay was used to verify the binding relationship between hsa-miR-450b-5p and lncRNA NEAT1. The expression levels of lncRNA NEAT1 and EZH2 in normal gastric epithelial cells (GES-1) and gastric cancer cells (MGC-803, SGC-7901 and MKN-28) were detected by qPCR and WB. MGC-803 and SGC-7901 cells were divided into the hsa-miR-450b-5p-mimic group, the mimic-NC group,the si-NEAT1 group and the si-NC group according to different transfections, and the overexpression and knockdown effects were verified by qPCR 36~48 h after transfection. qPCR and WB were used to detect and observe the effects of overexpression of hsa-miR-450b-5p on the protein expressions of lncRNA NEAT1 and EZH2mRNA, in cells, and the effect of knockdown of lncRNA NEAT1 on the expressions of hsa-miR-450b-5p and EZH2 mRNA. CCK-8 method, scratch healing assay and flow cytometry were used to detect the effects of knockdown of EZH2 or knockdown of lncRNA NEAT1 on cell proliferation, migration, and apoptosis, respectively. Results: The upstream miRNA and lncRNA of EZH2 obtained through bioinformatic analysis and screening were has-miR-450b-5p and lncRNA NEAT1, and the targeting relationship between the two was verified by double luciferase reporter gene assay. lncRNA NEAT1 and EZH2 mRNA 、protein were highly expressed in gastric cancer cells (both P<0.05). Compared with the mimic-NC group,the levels of miR-450b-5p in MGC-803 and SGC-7901 cells in the hsa-miR-450b-5p-mimic group increased significantly, while the expressions of EZH2 mRNA, protein and lncRNA NEAT1 decreased significantly (P<0.05 or P<0.01). Compared with the si-NC group, the expressions of lncRNA NEAT1 and EZH2 mRNA in MGC-803 and SGC-7901 cells decreased significantly in the si-NEAT1 group (both P<0.01), and the expression of hsa-miR-450b-5p in SGC-7901 cells increased significantly (P<0.05). The proliferation and migration abilities of MGC-803 and SGC-7901 cells were significantly reduced after knockdown of EZH2 or lncRNA NEAT1 (both P<0.01). Conclusion: lncRNA NEAT1 and EZH2 were highly expressed in gastric cancer cells, and lncRNA NEAT1 promoted the expression of EZH2 and improved the proliferation and migration abilities of gastric cancer MGC-803 and SGC-7901 cells through hsa-miR-450b-5p.
Objective: To screen the upstream miRNAs and lncRNAs of EZH2 gene, analyze their expressions in gastric cancer cells,verify the targeting relationship between them, and discuss their effects on the proliferation, migration and apoptosis of gastric cancer cells. Methods: The upstream miRNA (has-miR-450b-5p) of EZH2 was queried, analyzed and screened by ENCORI, miRDB and Target Scan databases, and the upstream lncRNA (lncRNA NEAT1) of hsa-miR-450b-5p was screened by ENCORI database and DAINA database. The binding sites between hsa-miR-450b-5p, lncRNA NEAT1 and EZH2 were predicted. Dual-luciferase reporter assay was used to verify the binding relationship between hsa-miR-450b-5p and lncRNA NEAT1. The expression levels of lncRNA NEAT1 and EZH2 in normal gastric epithelial cells (GES-1) and gastric cancer cells (MGC-803, SGC-7901 and MKN-28) were detected by qPCR and WB. MGC-803 and SGC-7901 cells were divided into the hsa-miR-450b-5p-mimic group, the mimic-NC group,the si-NEAT1 group and the si-NC group according to different transfections, and the overexpression and knockdown effects were verified by qPCR 36~48 h after transfection. qPCR and WB were used to detect and observe the effects of overexpression of hsa-miR-450b-5p on the protein expressions of lncRNA NEAT1 and EZH2mRNA, in cells, and the effect of knockdown of lncRNA NEAT1 on the expressions of hsa-miR-450b-5p and EZH2 mRNA. CCK-8 method, scratch healing assay and flow cytometry were used to detect the effects of knockdown of EZH2 or knockdown of lncRNA NEAT1 on cell proliferation, migration, and apoptosis, respectively. Results: The upstream miRNA and lncRNA of EZH2 obtained through bioinformatic analysis and screening were has-miR-450b-5p and lncRNA NEAT1, and the targeting relationship between the two was verified by double luciferase reporter gene assay. lncRNA NEAT1 and EZH2 mRNA 、protein were highly expressed in gastric cancer cells (both P<0.05). Compared with the mimic-NC group,the levels of miR-450b-5p in MGC-803 and SGC-7901 cells in the hsa-miR-450b-5p-mimic group increased significantly, while the expressions of EZH2 mRNA, protein and lncRNA NEAT1 decreased significantly (P<0.05 or P<0.01). Compared with the si-NC group, the expressions of lncRNA NEAT1 and EZH2 mRNA in MGC-803 and SGC-7901 cells decreased significantly in the si-NEAT1 group (both P<0.01), and the expression of hsa-miR-450b-5p in SGC-7901 cells increased significantly (P<0.05). The proliferation and migration abilities of MGC-803 and SGC-7901 cells were significantly reduced after knockdown of EZH2 or lncRNA NEAT1 (both P<0.01). Conclusion: lncRNA NEAT1 and EZH2 were highly expressed in gastric cancer cells, and lncRNA NEAT1 promoted the expression of EZH2 and improved the proliferation and migration abilities of gastric cancer MGC-803 and SGC-7901 cells through hsa-miR-450b-5p.
2024, 31(2):146-153.
DOI: 10.3872/j.issn.1007-385X.2024.02.005
Abstract:
To investigate the influence of atractylodin (ATR) on programmed death of non-small cell lung cancer (NSCLC) A549 cells and the growth of xenografts in nude mice by regulating receptor-interacting protein kinase (RIPK) 1/RIPK3/mixed lineage kinase domain like (MLKL) signaling pathway. Methods: A549 cells were treated with 0~160 μmol/L ATR, and the cell viability was detected by MTT method to determine the concentration of subsequent experiments. A549 cells were treated with ATR and/or RIPK1 inhibitor necrostatin-1 (Nec-1) and caspase inhibitor Z-VAD-FMK, to verify whether ATR induced programmed necrosis in A549 cells. A549 cells were divided into the control group, the ATR-L, ATR-M and ATR-H group (treated with 0, 10, 20 and 40 μmol/L ATR, respectively) and the ATR+Nec-1 group (treated with 40 μmol/L atractylodin and 50 μmol/L Nec-1). After 24 h of treatment, PI single staining and Hoechst33342/PI double staining were used to detect cell death; transmission electron microscopy (TEM) was used to observe the morphology of cell death; DCFH-DA fluorescent probe was used to detect intracellular ROS level; JC-1 staining was used to detect mitochondrial membrane potential, and WB method was used to detect the expression level of RIPK1/RIPK3/MLKL signaling pathway-related proteins in cells. A xenograft model of A549 cells was constructed in nude mice, and 10 mg/kg ATR (dissolved in corn oil) was administered to nude mice by gavage for 5 weeks to observe the effect of atractylodin on xenograft growth. The expression level of RIPK1/RIPK3/MLKL signaling pathway-related proteins in xenograft tissues was detected by WB method. Results: 10-160 μmol/L ATR could significantly inhibit the proliferation of A549 cells, and the concentrations of 10, 20 and 40 μmol/L were selected for follow-up experiments. The survival rate of A549 cells in the ATR group was significantly lower than that in the control group (P<0.01) and ATR+Nec-1 group (P<0.01), while the cell survival rate in the ATR+z-VAD group was significantly lower than that in the z-VAD group (P<0.01), indicating that ATR could induce programmed necrosis of A549 cells instead of apoptosis. Compared with the control group, A549 cells in the ATR-treated groups were swollen; the mitochondria were vacuolated; the inner ridge disappeared, the cell contents leaked outward, and the nuclei were aggregated, showing necrotic characteristics. The mortality rate, ROS level, expression levels of p-RIPK1, p-RIPK3 and p-MLKL in the ATR-L group, ATR-M group and ATR-H group A549 cells increased significantly, while the mitochondrial membrane potential decreased significantly (all P<0.01), all of which were concentration-dependent. Compared with the ATR-H group, the mortality rate, ROS level, and expression levels of p-RIPK1, p-RIPK3 and p-MLKL in the ATR+Nec-1 group decreased, while the mitochondrial membrane potential increased significantly (all P<0.01). The results of nude mouse xenograft experiment showed that compared with the control group, the volume and mass of xenografts were decreased (P<0.05 or P<0.01), and the protein expression levels of p-RIPK1, p-RIPK3 and p-MLKL in the tumor tissues in the ATR group increased significantly (all P<0.01). Conclusion: ATR may induce programmed necrosis of A549 cells by activating the RIPK1/RIPK3/MLKL signaling pathway, and inhibit the growth of A549 cells and their nude mouse xenografts.
To investigate the influence of atractylodin (ATR) on programmed death of non-small cell lung cancer (NSCLC) A549 cells and the growth of xenografts in nude mice by regulating receptor-interacting protein kinase (RIPK) 1/RIPK3/mixed lineage kinase domain like (MLKL) signaling pathway. Methods: A549 cells were treated with 0~160 μmol/L ATR, and the cell viability was detected by MTT method to determine the concentration of subsequent experiments. A549 cells were treated with ATR and/or RIPK1 inhibitor necrostatin-1 (Nec-1) and caspase inhibitor Z-VAD-FMK, to verify whether ATR induced programmed necrosis in A549 cells. A549 cells were divided into the control group, the ATR-L, ATR-M and ATR-H group (treated with 0, 10, 20 and 40 μmol/L ATR, respectively) and the ATR+Nec-1 group (treated with 40 μmol/L atractylodin and 50 μmol/L Nec-1). After 24 h of treatment, PI single staining and Hoechst33342/PI double staining were used to detect cell death; transmission electron microscopy (TEM) was used to observe the morphology of cell death; DCFH-DA fluorescent probe was used to detect intracellular ROS level; JC-1 staining was used to detect mitochondrial membrane potential, and WB method was used to detect the expression level of RIPK1/RIPK3/MLKL signaling pathway-related proteins in cells. A xenograft model of A549 cells was constructed in nude mice, and 10 mg/kg ATR (dissolved in corn oil) was administered to nude mice by gavage for 5 weeks to observe the effect of atractylodin on xenograft growth. The expression level of RIPK1/RIPK3/MLKL signaling pathway-related proteins in xenograft tissues was detected by WB method. Results: 10-160 μmol/L ATR could significantly inhibit the proliferation of A549 cells, and the concentrations of 10, 20 and 40 μmol/L were selected for follow-up experiments. The survival rate of A549 cells in the ATR group was significantly lower than that in the control group (P<0.01) and ATR+Nec-1 group (P<0.01), while the cell survival rate in the ATR+z-VAD group was significantly lower than that in the z-VAD group (P<0.01), indicating that ATR could induce programmed necrosis of A549 cells instead of apoptosis. Compared with the control group, A549 cells in the ATR-treated groups were swollen; the mitochondria were vacuolated; the inner ridge disappeared, the cell contents leaked outward, and the nuclei were aggregated, showing necrotic characteristics. The mortality rate, ROS level, expression levels of p-RIPK1, p-RIPK3 and p-MLKL in the ATR-L group, ATR-M group and ATR-H group A549 cells increased significantly, while the mitochondrial membrane potential decreased significantly (all P<0.01), all of which were concentration-dependent. Compared with the ATR-H group, the mortality rate, ROS level, and expression levels of p-RIPK1, p-RIPK3 and p-MLKL in the ATR+Nec-1 group decreased, while the mitochondrial membrane potential increased significantly (all P<0.01). The results of nude mouse xenograft experiment showed that compared with the control group, the volume and mass of xenografts were decreased (P<0.05 or P<0.01), and the protein expression levels of p-RIPK1, p-RIPK3 and p-MLKL in the tumor tissues in the ATR group increased significantly (all P<0.01). Conclusion: ATR may induce programmed necrosis of A549 cells by activating the RIPK1/RIPK3/MLKL signaling pathway, and inhibit the growth of A549 cells and their nude mouse xenografts.
2024, 31(2):154-160.
DOI: 10.3872/j.issn.1007-385X.2024.02.006
Abstract:
Objective: To investigate the effect of andrographolide (Andro) regulation of fatty acid synthase (Fas)/fatty acid synthase ligand (FasL) signaling axis on drug resistance of endometrial cancer Ishikawa cells. Methods: Ishikawa cells and cisplatin-resistant Ishikawa/DPP cells were treated respectively with 0, 5, 10 and 20 μg/mL DDP, and Ishikawa/DDP cells were treated with 0, 5, 10, 25 and 50 μmol/L Andro. Cell proliferation was detected by MTT and the appropriate dose of administration was selected for subsequent experiments. Ishikawa/DDP cells were randomly divided into the control group, the DDP group (intervention with DDP), the Andro group (intervention with DDP and Andro), the pcDNA3.1-NC group (transfection with pcDNA3.1 and intervention with DDP and Andro), and the pcDNA3.1-Fas/FasL group (transfection with pcDNA3.1-Fas/FasL and intervention with DDP and Andro). After 24 h of treatment, qPCR was used to detect the mRNA expression of Fas and FasL. Plate cloning assay; Transwell assay and flow cytometry were used to detect cell cloning ability, cell migration, invasion and apoptosis, respectively. The expressions of proliferating cell nuclear antigen (PCNA), BAX, Bcl-2, MMP-2, PD-L1, multidrug resistance protein-1 (MDR-1) and Fas and FasL proteins were detected by WB method. Results: DDP inhibited the proliferation of Ishikawa and Ishikawa/DPP cells in a dose-dependent manner. In addition, the sensitivity of Ishikawa/DPP cells to DDP was lower than that of Ishikawa cells (all P<0.05). Andro inhibited the proliferation of Ishikawa/DPP cells in a dose-dependent manner (all P<0.05). The expression levels of Fas and FasL in Ishikawa/DPP cells were higher than those in Ishikawa cells (all P<0.05). 20 μg/mL DDP and 25 μmol/L Andro were selected as the intervention doses, and the intervention time was 24 h. Compared with the control group, the mRNA and protein expression levels of PD-L1, MDR-1, Fas, FasL in Ishikawa/DPP cells in the DDP group increased significantly (P<0.05), but there were no significant differences in clone formation rate, number of migrating and invading cells and apoptosis rate (all P>0.05). Compared with the DDP group, the mRNA expression levels of Fas and FasL, the cell clone formation rate, the number of migrating and invading cells, the expression levl of PCNA, Bcl-2, MMP-2, PD-L1, MDR-1, Fas and FasL protein decreased significantly, while the expression level of BAX protein and the apoptosis rate increased significantly (P<0.05 or P<0.01). The pcDNA3.1-NC group was similar to the Andro group. Compared with the pcDNA3.1-NC group, the changes of Ishikawa/DPP cells in the pcDNA3.1-Fas/FasL group were reversed (P<0.05). Conclusion: Andro may inhibit the proliferation, migration and invasion of Ishikawa/DPP cells by inhibiting the Fas/FasL signaling axis, promote the apoptosis, and thereby reduce the resistance of cells to DDP.
Objective: To investigate the effect of andrographolide (Andro) regulation of fatty acid synthase (Fas)/fatty acid synthase ligand (FasL) signaling axis on drug resistance of endometrial cancer Ishikawa cells. Methods: Ishikawa cells and cisplatin-resistant Ishikawa/DPP cells were treated respectively with 0, 5, 10 and 20 μg/mL DDP, and Ishikawa/DDP cells were treated with 0, 5, 10, 25 and 50 μmol/L Andro. Cell proliferation was detected by MTT and the appropriate dose of administration was selected for subsequent experiments. Ishikawa/DDP cells were randomly divided into the control group, the DDP group (intervention with DDP), the Andro group (intervention with DDP and Andro), the pcDNA3.1-NC group (transfection with pcDNA3.1 and intervention with DDP and Andro), and the pcDNA3.1-Fas/FasL group (transfection with pcDNA3.1-Fas/FasL and intervention with DDP and Andro). After 24 h of treatment, qPCR was used to detect the mRNA expression of Fas and FasL. Plate cloning assay; Transwell assay and flow cytometry were used to detect cell cloning ability, cell migration, invasion and apoptosis, respectively. The expressions of proliferating cell nuclear antigen (PCNA), BAX, Bcl-2, MMP-2, PD-L1, multidrug resistance protein-1 (MDR-1) and Fas and FasL proteins were detected by WB method. Results: DDP inhibited the proliferation of Ishikawa and Ishikawa/DPP cells in a dose-dependent manner. In addition, the sensitivity of Ishikawa/DPP cells to DDP was lower than that of Ishikawa cells (all P<0.05). Andro inhibited the proliferation of Ishikawa/DPP cells in a dose-dependent manner (all P<0.05). The expression levels of Fas and FasL in Ishikawa/DPP cells were higher than those in Ishikawa cells (all P<0.05). 20 μg/mL DDP and 25 μmol/L Andro were selected as the intervention doses, and the intervention time was 24 h. Compared with the control group, the mRNA and protein expression levels of PD-L1, MDR-1, Fas, FasL in Ishikawa/DPP cells in the DDP group increased significantly (P<0.05), but there were no significant differences in clone formation rate, number of migrating and invading cells and apoptosis rate (all P>0.05). Compared with the DDP group, the mRNA expression levels of Fas and FasL, the cell clone formation rate, the number of migrating and invading cells, the expression levl of PCNA, Bcl-2, MMP-2, PD-L1, MDR-1, Fas and FasL protein decreased significantly, while the expression level of BAX protein and the apoptosis rate increased significantly (P<0.05 or P<0.01). The pcDNA3.1-NC group was similar to the Andro group. Compared with the pcDNA3.1-NC group, the changes of Ishikawa/DPP cells in the pcDNA3.1-Fas/FasL group were reversed (P<0.05). Conclusion: Andro may inhibit the proliferation, migration and invasion of Ishikawa/DPP cells by inhibiting the Fas/FasL signaling axis, promote the apoptosis, and thereby reduce the resistance of cells to DDP.
2024, 31(2):161-168.
DOI: 10.3872/j.issn.1007-385X.2024.02.007
Abstract:
Objective: To investigate the role of LINC00958/vascular endothelial growth factor C (VEGF-C) signaling pathway in lymphovascular formation and lymphatic metastasis in cervical cancer. Methods: 42 samples of cervical cancer tissues and corresponding adjacent tissues were collected from patients who underwent surgery at Henan Provincial People's Hospital between September 2020 and September 2022. The expressions of LINC00958 in cervical cancer specimens and cervical cancer cells (Hela, C33A, SiHa, Caski) were examined by qPCR. Caski cells were transfected with LINC00958 overexpression vector (LINC00958 group) or vehicle control (CMV group), and SiHa cells were transfected with shRNA sequences of knocked-down LINC00958 (shLINC00958 group) and VEGF-C (shVEGF-C group), or negative control shRNA (shNC group). The effects of overexpressed or knocked-down LINC00958 on the proliferation, migration and invasion of cervical cancer cells were detected by CCK-8 method and Transwell assay, respectively. The effect of the culture supernatant of transfected cells on the lymphovascular formation capacity of human lymphatic endothelial cells (HLEC) was observed. A mouse popliteal lymph node metastasis model was established to investigate the effects of overexpressed LINC00958 or simultaneous knockdown of VEGF-C on cervical cancer lymph node metastasis. Results: LINC00958 was highly expressed in cervical cancer tissues (P<0.001), and high levels of LINC00958 were associated with large tumors, advanced tumor grade, depth of invasion, and lymphatic metastasis (P<0.05 or P<0.01). Compared with that in the normal human cervical epithelial cells ende1617, the level of LINC00958 in cervical cancer cells was significantly increased (P<0.01 or P<0.001). The proliferation, migration and invasion abilities of SiHa cells in the shLINC00958 group and the HLEC lymph vessel pro-formation ability of the culture supernatant were significantly lower than those in the shNC group (P<0.05, P<0.01 or P<0.001), and the proliferation, migration and invasion abilities of Caski cells in the LINC00958 group and the HLEC lymph vessel pro-formation ability of the culture supernatant were significantly higher than those in the CMV group (P<0.05, P<0.01 or P<0.001). RNA pull-down and RNA immunoprecipitation assays showed that LINC00958 in cervical cancer cells could specifically bind to VEGF-C. The proliferation, migration and invasion abilities of Caski cells in the LINC00958+shVEGF-C group and the lymph vessel pro-formation ability of the culture supernatant were significantly lower than those in the LINC00958 group (P<0.01 or P<0.001). In the mouse popliteal lymph node metastasis model, the volume of popliteal lymph nodes in the LINC00958+shVEGF-C group and the proportion of VEGF-C protein, N-cadherin protein and LYVE-1 positive cells were significantly lower than those in the LINC00958 group (all P<0.001). Conclusion: LINC00958 directly interacts with VEGF-C protein to enhance the proliferation and invasion abilities and lymph vessel formation ability of cervical cancer cells, and promote lymph node metastasis in the mouse popliteal lymph node metastasis model.
Objective: To investigate the role of LINC00958/vascular endothelial growth factor C (VEGF-C) signaling pathway in lymphovascular formation and lymphatic metastasis in cervical cancer. Methods: 42 samples of cervical cancer tissues and corresponding adjacent tissues were collected from patients who underwent surgery at Henan Provincial People's Hospital between September 2020 and September 2022. The expressions of LINC00958 in cervical cancer specimens and cervical cancer cells (Hela, C33A, SiHa, Caski) were examined by qPCR. Caski cells were transfected with LINC00958 overexpression vector (LINC00958 group) or vehicle control (CMV group), and SiHa cells were transfected with shRNA sequences of knocked-down LINC00958 (shLINC00958 group) and VEGF-C (shVEGF-C group), or negative control shRNA (shNC group). The effects of overexpressed or knocked-down LINC00958 on the proliferation, migration and invasion of cervical cancer cells were detected by CCK-8 method and Transwell assay, respectively. The effect of the culture supernatant of transfected cells on the lymphovascular formation capacity of human lymphatic endothelial cells (HLEC) was observed. A mouse popliteal lymph node metastasis model was established to investigate the effects of overexpressed LINC00958 or simultaneous knockdown of VEGF-C on cervical cancer lymph node metastasis. Results: LINC00958 was highly expressed in cervical cancer tissues (P<0.001), and high levels of LINC00958 were associated with large tumors, advanced tumor grade, depth of invasion, and lymphatic metastasis (P<0.05 or P<0.01). Compared with that in the normal human cervical epithelial cells ende1617, the level of LINC00958 in cervical cancer cells was significantly increased (P<0.01 or P<0.001). The proliferation, migration and invasion abilities of SiHa cells in the shLINC00958 group and the HLEC lymph vessel pro-formation ability of the culture supernatant were significantly lower than those in the shNC group (P<0.05, P<0.01 or P<0.001), and the proliferation, migration and invasion abilities of Caski cells in the LINC00958 group and the HLEC lymph vessel pro-formation ability of the culture supernatant were significantly higher than those in the CMV group (P<0.05, P<0.01 or P<0.001). RNA pull-down and RNA immunoprecipitation assays showed that LINC00958 in cervical cancer cells could specifically bind to VEGF-C. The proliferation, migration and invasion abilities of Caski cells in the LINC00958+shVEGF-C group and the lymph vessel pro-formation ability of the culture supernatant were significantly lower than those in the LINC00958 group (P<0.01 or P<0.001). In the mouse popliteal lymph node metastasis model, the volume of popliteal lymph nodes in the LINC00958+shVEGF-C group and the proportion of VEGF-C protein, N-cadherin protein and LYVE-1 positive cells were significantly lower than those in the LINC00958 group (all P<0.001). Conclusion: LINC00958 directly interacts with VEGF-C protein to enhance the proliferation and invasion abilities and lymph vessel formation ability of cervical cancer cells, and promote lymph node metastasis in the mouse popliteal lymph node metastasis model.
2024, 31(2):169-174.
DOI: 10.3872/j.issn.1007-385X.2024.02.008
Abstract:
铜死亡是一种新发现的细胞程序性死亡方式，细胞内的铜代谢失衡尤其是铜离子过载有可能会导致细胞铜死亡的发生。过量的铜积累会靶向结合硫辛酰化三羧酸循环蛋白，使其异常聚集进而触发细胞死亡，铁氧化还原蛋白1（FDX1）等多个分子能够调控铜死亡活性。肿瘤细胞内铜代谢稳态主要靠四组铜相关蛋白质的相互作用来维持，影响受体酪氨酸激酶（RTK）、自噬、Notch 等相关信号通路，与肿瘤的发生发展息息相关。合理利用铜离子载体和铜螯合剂等铜配合物以及铜死亡相关生物标志物将有助于开发肿瘤治疗和预后评估新策略，在未来的肿瘤个性化治疗中有着巨大的潜力。
铜死亡是一种新发现的细胞程序性死亡方式，细胞内的铜代谢失衡尤其是铜离子过载有可能会导致细胞铜死亡的发生。过量的铜积累会靶向结合硫辛酰化三羧酸循环蛋白，使其异常聚集进而触发细胞死亡，铁氧化还原蛋白1（FDX1）等多个分子能够调控铜死亡活性。肿瘤细胞内铜代谢稳态主要靠四组铜相关蛋白质的相互作用来维持，影响受体酪氨酸激酶（RTK）、自噬、Notch 等相关信号通路，与肿瘤的发生发展息息相关。合理利用铜离子载体和铜螯合剂等铜配合物以及铜死亡相关生物标志物将有助于开发肿瘤治疗和预后评估新策略，在未来的肿瘤个性化治疗中有着巨大的潜力。
2024, 31(2):175-182.
DOI: 10.3872/j.issn.1007-385X.2024.02.009
Abstract:
PI3K-Akt 信号通路作为肿瘤相关十大信号通路之一，在调控肿瘤恶性进展中扮演着重要角色。PIK3CA编码PI3K复合体蛋白催化亚基P110α，是一种典型的致癌突变，对实体瘤的发生和发展至关重要。PIK3CA突变可导致肿瘤对一线抗癌药物产生耐药性，其机制可能与PIK3CA突变激活PI3K-Akt 信号通路相关。目前，几种靶向PIK3CA突变的抑制剂在临床研究中取得了一定进展，特别是PI3Kα特异性抑制剂阿培利司已被FDA批准作为PIK3CA突变乳腺癌的治疗药物。本文就PIK3CA突变促进肿瘤药物耐药的机制，以及逆转耐药的治疗策略作一综述，以期更全面了解PIK3CA突变在肿瘤耐药方面的进展以及最新治疗策略。
PI3K-Akt 信号通路作为肿瘤相关十大信号通路之一，在调控肿瘤恶性进展中扮演着重要角色。PIK3CA编码PI3K复合体蛋白催化亚基P110α，是一种典型的致癌突变，对实体瘤的发生和发展至关重要。PIK3CA突变可导致肿瘤对一线抗癌药物产生耐药性，其机制可能与PIK3CA突变激活PI3K-Akt 信号通路相关。目前，几种靶向PIK3CA突变的抑制剂在临床研究中取得了一定进展，特别是PI3Kα特异性抑制剂阿培利司已被FDA批准作为PIK3CA突变乳腺癌的治疗药物。本文就PIK3CA突变促进肿瘤药物耐药的机制，以及逆转耐药的治疗策略作一综述，以期更全面了解PIK3CA突变在肿瘤耐药方面的进展以及最新治疗策略。
2024, 31(2):183-188.
DOI: 10.3872/j.issn.1007-385X.2024.02.010
Abstract:
宫颈癌是全球女性第四大常见恶性肿瘤，由于其复杂性及异质性，患者病死率仍然较高。代谢重编程是肿瘤细胞的一个重要特征，通过癌基因的突变、抑癌基因的失活、信号转导途径失调和肿瘤微环境（TME）紊乱等发挥代谢重编程的致癌作用。随着从人乳头状瘤病毒（HPV）感染到宫颈癌发生至侵袭与转移，宫颈癌代谢表型也在演变。HPV癌蛋白通过诱导代谢相关基因的表达改变细胞代谢模式促进宫颈癌的发生，上调有氧糖酵解以满足肿瘤细胞的增殖，增强脂肪酸代谢和有氧糖酵解以满足宫颈癌的淋巴结转移。此外，代谢重编程通过多种机制影响宫颈癌的治疗，例如代谢产物改变导致氧化应激失调介导的顺铂耐药、代谢相关基因重塑TME而出现的免疫抑制。在某些情况下，针对这些代谢途径和相关代谢酶的代谢抑制剂联合免疫治疗可成为宫颈癌的新型治疗方式。本文综述了代谢重编程在宫颈癌中的研究进展，为研发新的生物标志物和治疗靶点提供了新的策略。
宫颈癌是全球女性第四大常见恶性肿瘤，由于其复杂性及异质性，患者病死率仍然较高。代谢重编程是肿瘤细胞的一个重要特征，通过癌基因的突变、抑癌基因的失活、信号转导途径失调和肿瘤微环境（TME）紊乱等发挥代谢重编程的致癌作用。随着从人乳头状瘤病毒（HPV）感染到宫颈癌发生至侵袭与转移，宫颈癌代谢表型也在演变。HPV癌蛋白通过诱导代谢相关基因的表达改变细胞代谢模式促进宫颈癌的发生，上调有氧糖酵解以满足肿瘤细胞的增殖，增强脂肪酸代谢和有氧糖酵解以满足宫颈癌的淋巴结转移。此外，代谢重编程通过多种机制影响宫颈癌的治疗，例如代谢产物改变导致氧化应激失调介导的顺铂耐药、代谢相关基因重塑TME而出现的免疫抑制。在某些情况下，针对这些代谢途径和相关代谢酶的代谢抑制剂联合免疫治疗可成为宫颈癌的新型治疗方式。本文综述了代谢重编程在宫颈癌中的研究进展，为研发新的生物标志物和治疗靶点提供了新的策略。
2024, 31(2):189-195.
DOI: 10.3872/j.issn.1007-385X.2024.02.011
Abstract:
靶向治疗和免疫治疗常作为晚期肝细胞癌的系统性抗肿瘤方案。然而，受到多种因素的影响，如肿瘤微环境、细胞信号的转导通路、药物转运等因素的影响，晚期肝细胞癌的患者在治疗时往往出现不同程度的原发性和获得性耐药，患者长期获益有限。自IMbrave150 试验发现靶向联合免疫治疗方案优于单药治疗后，有效改善了过去单药治疗耐药所产生的生存获益受限，而获得性耐药的解决方案仍受限于缺乏统一标准、病例难收集、差异基因较少等因素。本文综述目前众多正在进行的临床试验，希望为克服肝细胞癌耐药问题提供参考。
靶向治疗和免疫治疗常作为晚期肝细胞癌的系统性抗肿瘤方案。然而，受到多种因素的影响，如肿瘤微环境、细胞信号的转导通路、药物转运等因素的影响，晚期肝细胞癌的患者在治疗时往往出现不同程度的原发性和获得性耐药，患者长期获益有限。自IMbrave150 试验发现靶向联合免疫治疗方案优于单药治疗后，有效改善了过去单药治疗耐药所产生的生存获益受限，而获得性耐药的解决方案仍受限于缺乏统一标准、病例难收集、差异基因较少等因素。本文综述目前众多正在进行的临床试验，希望为克服肝细胞癌耐药问题提供参考。
2024, 31(2):196-200.
DOI: 10.3872/j.issn.1007-385X.2024.02.012
Abstract:
P66Shc 作为ShcA家族成员，具有家族特有的高度保守结构域，对细胞的增殖、分化和凋亡起重要的调控作用，因而在肿瘤发生发展中也起着关键性作用。在不同的肿瘤细胞中，p66Shc 的表达表现出两面性：既可促进肿瘤生长和转移，也可以抑制肿瘤发展。异常水平表达的p66Shc 通常与肿瘤细胞过度增殖、高转移风险和不良预后相关。P66Shc 还可通过激活氧化应激通路来调节肿瘤细胞的代谢状态，参与协调肿瘤细胞凋亡、自噬和失巢凋亡等不同死亡方式。探究p66Shc 在肿瘤发生发展中的作用及机制，可为肿瘤临床治疗中有效靶点的寻找带来更多新的突破。
P66Shc 作为ShcA家族成员，具有家族特有的高度保守结构域，对细胞的增殖、分化和凋亡起重要的调控作用，因而在肿瘤发生发展中也起着关键性作用。在不同的肿瘤细胞中，p66Shc 的表达表现出两面性：既可促进肿瘤生长和转移，也可以抑制肿瘤发展。异常水平表达的p66Shc 通常与肿瘤细胞过度增殖、高转移风险和不良预后相关。P66Shc 还可通过激活氧化应激通路来调节肿瘤细胞的代谢状态，参与协调肿瘤细胞凋亡、自噬和失巢凋亡等不同死亡方式。探究p66Shc 在肿瘤发生发展中的作用及机制，可为肿瘤临床治疗中有效靶点的寻找带来更多新的突破。
2024, 31(2):201-206.
DOI: 10.3872/j.issn.1007-385X.2024.02.013
Abstract:
免疫检查点抑制剂（ICI）是肿瘤治疗的革命性突破，在治疗肿瘤的同时，可以引起各系统的免疫相关不良反应（irAE）的发生。本文报道2例恶性肿瘤晚期患者接受ICI 治疗后出现TEN 的病例，分别为1例早期SJS/TEN 病例和1例难治性TEN 病例，两例患者均应用血浆置换治疗作为一线治疗，其临床症状及生活质量功能均获得改善，结合系统性内科治疗，2例患者后期均获得痊愈且无复发。ICI 相关TEN发生率低，但死亡率高，故提高对该病的早期识别及及时救治至关重要。此2病例早期应用血浆置换治疗效果较好，为ICI相关重度irAE的临床诊疗提供参考。
免疫检查点抑制剂（ICI）是肿瘤治疗的革命性突破，在治疗肿瘤的同时，可以引起各系统的免疫相关不良反应（irAE）的发生。本文报道2例恶性肿瘤晚期患者接受ICI 治疗后出现TEN 的病例，分别为1例早期SJS/TEN 病例和1例难治性TEN 病例，两例患者均应用血浆置换治疗作为一线治疗，其临床症状及生活质量功能均获得改善，结合系统性内科治疗，2例患者后期均获得痊愈且无复发。ICI 相关TEN发生率低，但死亡率高，故提高对该病的早期识别及及时救治至关重要。此2病例早期应用血浆置换治疗效果较好，为ICI相关重度irAE的临床诊疗提供参考。
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
您是第位访问者
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.