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Volume 31,Issue 7,2024 Table of Contents
2024, 31(7):647-654.
DOI: 10.3872/j.issn.1007-385X.2024.07.001
Abstract:
Chimeric antigen receptor gene-modified T (CAR-T) cell immunotherapy is considered as one of the most promising tumor treatments. The number of effector CAR-T cells is a key factor in determining the therapeutic effect of CAR-T cell therapy. The expansion of CAR-T cells in vitro is time and energy-consuming. After transfusion, it’s difficult for CAR-T cells to infiltrate into solid tumors, resulting in a significant decrease in the number of CAR-T cells that can effectively inhibit solid tumors. Currently, the amplification of CAR-T cells has problems in enhancing amplification specificity and treatment safety, which hinders the clinical transformation of CAR-T cell therapy. In recent years, achievements in new immunity agonist and their downstream signals have provided more options for CAR-T amplification, and the novel administration methods of immunity agonist have further improved the safety of CAR-T amplification in vivo. This review analyzes the current challenges in CAR-T cell amplification, and systematically expounds the new strategies of CAR-T cell amplification in vitro and in vivo in recent years, providing new thoughts for the efficacy and capacity optimization of CAR-T cell therapy.
Chimeric antigen receptor gene-modified T (CAR-T) cell immunotherapy is considered as one of the most promising tumor treatments. The number of effector CAR-T cells is a key factor in determining the therapeutic effect of CAR-T cell therapy. The expansion of CAR-T cells in vitro is time and energy-consuming. After transfusion, it’s difficult for CAR-T cells to infiltrate into solid tumors, resulting in a significant decrease in the number of CAR-T cells that can effectively inhibit solid tumors. Currently, the amplification of CAR-T cells has problems in enhancing amplification specificity and treatment safety, which hinders the clinical transformation of CAR-T cell therapy. In recent years, achievements in new immunity agonist and their downstream signals have provided more options for CAR-T amplification, and the novel administration methods of immunity agonist have further improved the safety of CAR-T amplification in vivo. This review analyzes the current challenges in CAR-T cell amplification, and systematically expounds the new strategies of CAR-T cell amplification in vitro and in vivo in recent years, providing new thoughts for the efficacy and capacity optimization of CAR-T cell therapy.
2 Expression of PD-1 shRNA enhances the killing ability of CD19-targeting CAR-T cells on tumor cells
2024, 31(7):655-661.
DOI: 10.3872/j.issn.1007-385X.2024.07.002
Abstract:
Objective: To design and construct CD19-targeting CAR-T cells expressing PD-1 shRNA and validate their anti-tumor function in vitro. Methods: The authors designed and constructed CD19 CAR molecule gene expressing PD-1 shRNA, and packaged them into retroviral vector using packaging cells. The viral vector copy number was detected by qPCR, and then human primary T cells were transduced to obtain CAR-T cells, which were divided into three groups: RNAU6-CD19 CAR-T, PD-1 shRNA1-CD19 CAR-T, and PD-1 shRNA2- CD19 CAR-T cells. qPCR was applied to detect the expression levels of PD-1 mRNA in three groups of CAR-T cells. Flow cytometry was used to detect the expression level of PD-1 on CAR-T cells in three groups. The luciferase reporter gene method and flow cytometry were used to detect the killing function of CAR-T cells against target cells (human lymphoma daudi cells) at different efficacy to target ratios. Results: Three groups of CAR molecules, namely RNAU6-CD19 CAR, PD-1 shRNA1-CD19 CAR and PD-1 shRNA2-CD19 CAR, were successfully packaged into retroviral vector, in which all retroviral vector copy numbers were higher than 1×107 copies/mL. CAR-T cells were obtained by transducing human primary T cells. The CAR-T transduction efficiencies of RNAU6-CD19 CAR-T, PD-1 shRNA1- CD19 CAR-T and PD-1 shRNA2-CD19 CAR-T cells were 43.1%, 55.1%, and 41.7% respectively. Compared with RNAU6-CD19 CAR-T cells, PD-1 shRNA1-CD19 CAR-T and PD-1 shRNA2-CD19 CAR-T cells showed a significant decrease in the expression level of PD-1 mRNA (all PPin vitro (PPConclusion: Successfull construction of CD19-targeting CAR-T cells expressing PD-1 shRNA can improve the killing efficiency against CD19 positive target cells, reduce the expressions of PD-1 mRNA and its translation product PD-1, and slow down the depletion of CAR-T cells.
Objective: To design and construct CD19-targeting CAR-T cells expressing PD-1 shRNA and validate their anti-tumor function in vitro. Methods: The authors designed and constructed CD19 CAR molecule gene expressing PD-1 shRNA, and packaged them into retroviral vector using packaging cells. The viral vector copy number was detected by qPCR, and then human primary T cells were transduced to obtain CAR-T cells, which were divided into three groups: RNAU6-CD19 CAR-T, PD-1 shRNA1-CD19 CAR-T, and PD-1 shRNA2- CD19 CAR-T cells. qPCR was applied to detect the expression levels of PD-1 mRNA in three groups of CAR-T cells. Flow cytometry was used to detect the expression level of PD-1 on CAR-T cells in three groups. The luciferase reporter gene method and flow cytometry were used to detect the killing function of CAR-T cells against target cells (human lymphoma daudi cells) at different efficacy to target ratios. Results: Three groups of CAR molecules, namely RNAU6-CD19 CAR, PD-1 shRNA1-CD19 CAR and PD-1 shRNA2-CD19 CAR, were successfully packaged into retroviral vector, in which all retroviral vector copy numbers were higher than 1×107 copies/mL. CAR-T cells were obtained by transducing human primary T cells. The CAR-T transduction efficiencies of RNAU6-CD19 CAR-T, PD-1 shRNA1- CD19 CAR-T and PD-1 shRNA2-CD19 CAR-T cells were 43.1%, 55.1%, and 41.7% respectively. Compared with RNAU6-CD19 CAR-T cells, PD-1 shRNA1-CD19 CAR-T and PD-1 shRNA2-CD19 CAR-T cells showed a significant decrease in the expression level of PD-1 mRNA (all PPin vitro (PPConclusion: Successfull construction of CD19-targeting CAR-T cells expressing PD-1 shRNA can improve the killing efficiency against CD19 positive target cells, reduce the expressions of PD-1 mRNA and its translation product PD-1, and slow down the depletion of CAR-T cells.
3 DKK1 blockade improves anti-tumor immune response by promoting Th1 type polarization of CD4+ T cells
2024, 31(7):662-668.
DOI: 10.3872/j.issn.1007-385X.2024.07.003
Abstract:
Objective: To explore the effect of Dickkopf-1 (DKK1) expression on CD4+ T cell polarization in malignant tumors and its potential value as a target for cancer immunotherapy. Methods: Bioinformatics algorithm was used to analyze the expression of DKK1 in multiple types of tumor tissues and adjacent tissues, and the correlation between the expression of DKK1 and the prognosis of cancer patients and immune infiltration of tumor microenvironment. The effect of DKK1 protein on the phenotype of CD4+ T cells was analyzed by flow cytometry in vitro. A melanoma B16F10 cell mouse subcutaneous transplantation tumor model was established to observe the effects of blocking DKK1 on the growth of mouse transplantation tumor and immune cell infiltration and phenotype in transplantation tumor tissues. Results: The mRNA expression levels of DKK1 in many kinds of tumor tissues were significantly higher than those in adjacent tissues. High expression of DKK1 was related to the poor prognosis of most tumor patients. In most types of tumors DKK1 played important negative regulatory role in CD4+ T cell anti-tumor immune response（ PP<0.01）. The results of flow cytometry in vitro showed that DKK1 protein stimulation significantly decreased the expression levels of T-bet, IFN-γ and CD107a in CD4+ T cells (all PP+ CD4+ ) increasing significantly (P+ T cells (CD44+ CD62L- ) increasing significantly (P+ CD4+ ) and Treg cells decreasing significantly (both PConclusion: Blocking DKK1 can effectively promote the phenotypic polarization of CD4+ T cells to Th1. DKK1 has potential value as a target for tumor immunotherapy.
Objective: To explore the effect of Dickkopf-1 (DKK1) expression on CD4+ T cell polarization in malignant tumors and its potential value as a target for cancer immunotherapy. Methods: Bioinformatics algorithm was used to analyze the expression of DKK1 in multiple types of tumor tissues and adjacent tissues, and the correlation between the expression of DKK1 and the prognosis of cancer patients and immune infiltration of tumor microenvironment. The effect of DKK1 protein on the phenotype of CD4+ T cells was analyzed by flow cytometry in vitro. A melanoma B16F10 cell mouse subcutaneous transplantation tumor model was established to observe the effects of blocking DKK1 on the growth of mouse transplantation tumor and immune cell infiltration and phenotype in transplantation tumor tissues. Results: The mRNA expression levels of DKK1 in many kinds of tumor tissues were significantly higher than those in adjacent tissues. High expression of DKK1 was related to the poor prognosis of most tumor patients. In most types of tumors DKK1 played important negative regulatory role in CD4+ T cell anti-tumor immune response（ PP<0.01）. The results of flow cytometry in vitro showed that DKK1 protein stimulation significantly decreased the expression levels of T-bet, IFN-γ and CD107a in CD4+ T cells (all PP+ CD4+ ) increasing significantly (P+ T cells (CD44+ CD62L- ) increasing significantly (P+ CD4+ ) and Treg cells decreasing significantly (both PConclusion: Blocking DKK1 can effectively promote the phenotypic polarization of CD4+ T cells to Th1. DKK1 has potential value as a target for tumor immunotherapy.
2024, 31(7):669-674.
DOI: 10.3872/j.issn.1007-385X.2024.07.004
Abstract:
Objective: To express the glycolytic enzyme alpha-enolase (ENO1) and its three enzyme active site deletion mutant ENO1 proteins (ENO-M1, pFastBac-M2, ENO1-M3) using a baculovirus expression vector system (insect BEVS), laying the groundwork for the subsequent study of metabolic therapy for cervical cancers. Methods: Molecule cloning was used to insert optimized ENO1 gene sequence into the pFastBacTM1 vector to obtain the recombinant plasmid pFastBac-ENO1 with target genes. Three active sites essential for ENO1's glycolytic function were deleted, and the corresponding optimized sequences were inserted into the pFastBacTM1 vector to generate the recombinant plasmids with three active site deletion, namely pFastBac-M1, pFastBac-M2 and pFastBac-M3. Recombinant baculoviruses rBV-ENO1, rBV-M1, rBV-M2 and rBV-M3 were subsequently obtained through transposition and transfection. The expression and specificity of the target proteins were examined using WB assay. Results: Recombinant bacilli rBacmid-ENO1, rBacmid-M1, rBacmid-M2 and rBacmid-M3 were successfully amplified, obtaining a gene fragment of about 2 000 bp in size, which was consistent with the expected size. The insect BEVS could express the ENO1 protein and its recombinant proteins (ENO1-M1, ENO1-M2, ENO1-M3) with three enzyme active site deletions, each with a molecular weight of approximately 52 000, as expected. WB analysis confirmed that these proteins reacted with the specific His-tag antibody. Conclusion: The insect BEV successfully expresses the target protein and its proteins with enzyme active site deletions, namely ENO1-M1, ENO1-M2 and ENO1-M3. This protein's reactivity establishes the foundation for subsequent determination of the affinity of these proteins, and ENO1 monoclonal antibody.
Objective: To express the glycolytic enzyme alpha-enolase (ENO1) and its three enzyme active site deletion mutant ENO1 proteins (ENO-M1, pFastBac-M2, ENO1-M3) using a baculovirus expression vector system (insect BEVS), laying the groundwork for the subsequent study of metabolic therapy for cervical cancers. Methods: Molecule cloning was used to insert optimized ENO1 gene sequence into the pFastBacTM1 vector to obtain the recombinant plasmid pFastBac-ENO1 with target genes. Three active sites essential for ENO1's glycolytic function were deleted, and the corresponding optimized sequences were inserted into the pFastBacTM1 vector to generate the recombinant plasmids with three active site deletion, namely pFastBac-M1, pFastBac-M2 and pFastBac-M3. Recombinant baculoviruses rBV-ENO1, rBV-M1, rBV-M2 and rBV-M3 were subsequently obtained through transposition and transfection. The expression and specificity of the target proteins were examined using WB assay. Results: Recombinant bacilli rBacmid-ENO1, rBacmid-M1, rBacmid-M2 and rBacmid-M3 were successfully amplified, obtaining a gene fragment of about 2 000 bp in size, which was consistent with the expected size. The insect BEVS could express the ENO1 protein and its recombinant proteins (ENO1-M1, ENO1-M2, ENO1-M3) with three enzyme active site deletions, each with a molecular weight of approximately 52 000, as expected. WB analysis confirmed that these proteins reacted with the specific His-tag antibody. Conclusion: The insect BEV successfully expresses the target protein and its proteins with enzyme active site deletions, namely ENO1-M1, ENO1-M2 and ENO1-M3. This protein's reactivity establishes the foundation for subsequent determination of the affinity of these proteins, and ENO1 monoclonal antibody.
2024, 31(7):675-680.
DOI: 10.3872/j.issn.1007-385X.2024.07.005
Abstract:
Objective: To investigate whether galangin (Gal) affects the proliferation, migration, invasion and apoptosis of osteosarcoma MG63 cells by regulating the cGAS/STING signaling pathway. Methods: Human osteosarcoma MG63 cells were cultured in vitro and treated with Gal at concentrations of 0, 5, 15, 25, 50, 100 and 200 μmol/L for 48 hours, and the effect of Gal on cell viability was detected by CCK-8 method. MG63 cells were divided into the control group (untreated cells), the Gal low concentration group (Gal-L group, treated with 50 μmol/L Gal), the Gal high concentration group (Gal-H group, treated with 100 μmol/L Gal), and the Gal-H+STING inhibitor group (Gal-H+H-151 group, treated with 100 μmol/L Gal+8 μmol/L H-151). EdU staining method, scratch healing test, Transwell chamber method and flow cytometry were used to detect cell proliferation, migration, invasion and apoptosis of cells in each group. Western blot was applied to detect the expression levels of cGAS and STING proteins in cells of each group. Results: Compared with the control group, the proliferation activity, migration, and invasion abilities of the cells in the Gal-L and Gal-H groups were significantly decreased (all PPPPConclusion: Gal may inhibit the proliferation, migration and invasion and promote the apoptosis of osteosarcoma cells by activating the cGAS/STING signaling pathway.
Objective: To investigate whether galangin (Gal) affects the proliferation, migration, invasion and apoptosis of osteosarcoma MG63 cells by regulating the cGAS/STING signaling pathway. Methods: Human osteosarcoma MG63 cells were cultured in vitro and treated with Gal at concentrations of 0, 5, 15, 25, 50, 100 and 200 μmol/L for 48 hours, and the effect of Gal on cell viability was detected by CCK-8 method. MG63 cells were divided into the control group (untreated cells), the Gal low concentration group (Gal-L group, treated with 50 μmol/L Gal), the Gal high concentration group (Gal-H group, treated with 100 μmol/L Gal), and the Gal-H+STING inhibitor group (Gal-H+H-151 group, treated with 100 μmol/L Gal+8 μmol/L H-151). EdU staining method, scratch healing test, Transwell chamber method and flow cytometry were used to detect cell proliferation, migration, invasion and apoptosis of cells in each group. Western blot was applied to detect the expression levels of cGAS and STING proteins in cells of each group. Results: Compared with the control group, the proliferation activity, migration, and invasion abilities of the cells in the Gal-L and Gal-H groups were significantly decreased (all PPPPConclusion: Gal may inhibit the proliferation, migration and invasion and promote the apoptosis of osteosarcoma cells by activating the cGAS/STING signaling pathway.
2024, 31(7):681-686.
DOI: 10.3872/j.issn.1007-385X.2024.07.006
Abstract:
Objective: To explore the effects of UCP2 inhibitor genipin (GEN) on the proliferation and mitochondrial function of human hypopharyngeal carcinoma FaDu cells. Methods: FaDu cells were treated with different concentrations of GEN for 24 hours and divided into the GEN 0 μmol/L (control) group, the 50 μmol/L group, the 100 μmol/L group, the 200 μmol/L group and the 400 μmol/L group. The CCK-8 method was employed to assess cell proliferation, and the DCFH-DA probe and JC-1 flow cytometry were utilized to measure the impact of GEN on reactive oxygen species (ROS) levels and mitochondrial membrane potential in FaDu cells. Laser confocal microscopy was utilized to observe the effect of GEN on the mitochondrial membrane permeability transition pore (MPTP) in FaDu cells. Spectrophotometry was employed to measure lactate levels in cells, and Western blot analysis was conducted to monitor changes in UCP2 protein expression. Results: In comparison with the control group, GEN significantly inhibited the proliferation activity of FaDu cells (PPPPPPPPConclusion: GEN modulates the expression of UCP2 in cells, consequently altering their redox potential and mitochondrial function, thus inhibiting the viability and inducing apoptosis in human nasopharyngeal carcinoma FaDu cells.
Objective: To explore the effects of UCP2 inhibitor genipin (GEN) on the proliferation and mitochondrial function of human hypopharyngeal carcinoma FaDu cells. Methods: FaDu cells were treated with different concentrations of GEN for 24 hours and divided into the GEN 0 μmol/L (control) group, the 50 μmol/L group, the 100 μmol/L group, the 200 μmol/L group and the 400 μmol/L group. The CCK-8 method was employed to assess cell proliferation, and the DCFH-DA probe and JC-1 flow cytometry were utilized to measure the impact of GEN on reactive oxygen species (ROS) levels and mitochondrial membrane potential in FaDu cells. Laser confocal microscopy was utilized to observe the effect of GEN on the mitochondrial membrane permeability transition pore (MPTP) in FaDu cells. Spectrophotometry was employed to measure lactate levels in cells, and Western blot analysis was conducted to monitor changes in UCP2 protein expression. Results: In comparison with the control group, GEN significantly inhibited the proliferation activity of FaDu cells (PPPPPPPPConclusion: GEN modulates the expression of UCP2 in cells, consequently altering their redox potential and mitochondrial function, thus inhibiting the viability and inducing apoptosis in human nasopharyngeal carcinoma FaDu cells.
2024, 31(7):687-693.
DOI: 10.3872/j.issn.1007-385X.2024.07.007
Abstract:
Objective: To investigate the effects of pristimerin (Pris) on the proliferation, apoptosis and vasculogenic mimicry (VM) of cervical cancer HeLa cells by regulating the Shh/Gli1 signaling pathway. Methods: MTT method was used to detect the inhibitory effects of Pris in different concentrations on the proliferation of cervical-cancer HeLa cells to select appropriate intervention concentration. Cervical cancer HeLa cells were grouped into the control group, the cyclopamine group, the Pris group, the Pris+pc-NC group and the Pris+pc-Shh group. MTT and EdU were applied to detect the proliferation abilities of cells in each group. Transwell chamber method and flow cytometry were used to detect cell migration and invasion abilities and cell apoptosis rate. In vitro angiogenesis experiments were used to observe the formation of VM. qPCR was used to detect Shh and Gli1 mRNA expression levels in each group. Western blotting was used to detect the expression levels of vascular endothelial growth factor A (VEGF-A), vascular endothelial cadherin (VE-cadherin), Ki-67, caspase-3, and proteins related to the Shh/Gli1 signaling pathway. Results: 0.25-2.5 mol/L Pris significantly inhibited the proliferation of HeLa cells, and 1.5 mol/L was selected for subsequent experiments. The cells in the control group formed a good lumen structure. Compared with the control group, the lumen structures of HeLa cells in the cyclopamine group, the Pris group and the Pris+pc-NC group were significantly damaged. Cell proliferation activity, proliferation rate, numbers of migration and invasion, expression levels of Shh and Gli1 mRNA, and expression levels of VEGF-A, VE-cadherin, Ki-67, Shh and Gli1 proteins were significantly decreased (all P<0.05). The apoptosis rate and the expression level of caspase-3 were significantly increased (all P<0.05). There was no significant difference in HeLa cell detection indicators between the cyclopamine group and the Pris group (all P>0.05). Compared with the Pris+pc-NC group, the formation of cell lumen structure in the Pris+pc-Shh group was significantly improved. The cell proliferation activity and proliferation rate, the numbers of migration and invasion cells, the expressions of Shh and Gli1 mRNA, and the expressions of VEGF-A, VE-cadherin, Ki-67, Shh and Gli1 proteins were significantly increased (all P<0.05). The apoptosis rate and the expression level of caspase-3 were significantly decreased (all P<0.05). Conclusion: Pris can inhibit the proliferation, migration, invasion and VM formation of cervical cancer HeLa cells, and promote cell apoptosis, which may be related to blocking the Shh/Gli1 signaling pathway.
Objective: To investigate the effects of pristimerin (Pris) on the proliferation, apoptosis and vasculogenic mimicry (VM) of cervical cancer HeLa cells by regulating the Shh/Gli1 signaling pathway. Methods: MTT method was used to detect the inhibitory effects of Pris in different concentrations on the proliferation of cervical-cancer HeLa cells to select appropriate intervention concentration. Cervical cancer HeLa cells were grouped into the control group, the cyclopamine group, the Pris group, the Pris+pc-NC group and the Pris+pc-Shh group. MTT and EdU were applied to detect the proliferation abilities of cells in each group. Transwell chamber method and flow cytometry were used to detect cell migration and invasion abilities and cell apoptosis rate. In vitro angiogenesis experiments were used to observe the formation of VM. qPCR was used to detect Shh and Gli1 mRNA expression levels in each group. Western blotting was used to detect the expression levels of vascular endothelial growth factor A (VEGF-A), vascular endothelial cadherin (VE-cadherin), Ki-67, caspase-3, and proteins related to the Shh/Gli1 signaling pathway. Results: 0.25-2.5 mol/L Pris significantly inhibited the proliferation of HeLa cells, and 1.5 mol/L was selected for subsequent experiments. The cells in the control group formed a good lumen structure. Compared with the control group, the lumen structures of HeLa cells in the cyclopamine group, the Pris group and the Pris+pc-NC group were significantly damaged. Cell proliferation activity, proliferation rate, numbers of migration and invasion, expression levels of Shh and Gli1 mRNA, and expression levels of VEGF-A, VE-cadherin, Ki-67, Shh and Gli1 proteins were significantly decreased (all P<0.05). The apoptosis rate and the expression level of caspase-3 were significantly increased (all P<0.05). There was no significant difference in HeLa cell detection indicators between the cyclopamine group and the Pris group (all P>0.05). Compared with the Pris+pc-NC group, the formation of cell lumen structure in the Pris+pc-Shh group was significantly improved. The cell proliferation activity and proliferation rate, the numbers of migration and invasion cells, the expressions of Shh and Gli1 mRNA, and the expressions of VEGF-A, VE-cadherin, Ki-67, Shh and Gli1 proteins were significantly increased (all P<0.05). The apoptosis rate and the expression level of caspase-3 were significantly decreased (all P<0.05). Conclusion: Pris can inhibit the proliferation, migration, invasion and VM formation of cervical cancer HeLa cells, and promote cell apoptosis, which may be related to blocking the Shh/Gli1 signaling pathway.
2024, 31(7):694-699.
DOI: 10.3872/j.issn.1007-385X.2024.07.008
Abstract:
Objective: To investigate the effects of isoliensinine (Iso) on the proliferation, apoptosis and autophagy of colon cancer SW480 cells through PI3K/Akt/mTOR signaling pathway. Methods: Colon cancer SW480 cells were treated with 10, 20 and 40 μmol/L Iso, and the effects of Iso on the cell proliferation capacity, apoptosis and expressions of autophagy related proteins LC3Ⅰ, LC3Ⅱ and p62 were detected by CCK-8, flow cytometry and Western blot, respectively. Then, SW480 cells were treated respectively with 20 μmol/L Iso and 25 μmol/L PI3K activator 740 Y-P, and the cells were divided into the control group, the 740 Y-P group, the Iso group and the Iso+740 Y-P group. The effects of 740 Y-P on the apoptosis and the expressions of LC3Ⅰ, LC3Ⅱ, p62, PI3K, p-PI3K, mTOR and p-mTOR proteins in each group were detected by flow cytometry and WB. Results: After treatments with 10, 20 and 40 μmol/L Iso, the proliferation capacity of SW480 cells was decreased significantly (all P<0.05); the apoptosis rate was increased significantly (all P<0.05); , the expressions of LC3Ⅱ/LC3Ⅰwere up-regulated significantly (all P<0.05), and the expression of p26 protein was down-regulated significantly (all P<0.05). After treatments with Iso and 740 Y-P, compared with the control group, the apoptosis rate and LC3Ⅱ/LC3Ⅰexpression of the 740 Y-P group were decreased significantly (both P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were increased significantly (all P<0.05).The apoptosis rate and LC3Ⅱ/LC3 expression in the Iso group were increased (both P<0.05) and the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the 740 Y-P group, the apoptosis rate and LC3Ⅱ/LC3Ⅰexpression were increased in the Iso+740 Y-P group (P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the Iso group, the apoptosis rate and LC3Ⅱ/LC3Ⅰexpression were decreased (both P<0.05), and the expressions of p26, p-PI3K/PI3K, and p-mTOR/mTOR were increased significantly (all P<0.05) in the Iso+740 Y-P group. Conclusion: Iso inhibits the proliferation and induces the apoptosis and autophagy of SW480 cells by inhibiting PI3K/Akt/mTOR signaling pathway.
Objective: To investigate the effects of isoliensinine (Iso) on the proliferation, apoptosis and autophagy of colon cancer SW480 cells through PI3K/Akt/mTOR signaling pathway. Methods: Colon cancer SW480 cells were treated with 10, 20 and 40 μmol/L Iso, and the effects of Iso on the cell proliferation capacity, apoptosis and expressions of autophagy related proteins LC3Ⅰ, LC3Ⅱ and p62 were detected by CCK-8, flow cytometry and Western blot, respectively. Then, SW480 cells were treated respectively with 20 μmol/L Iso and 25 μmol/L PI3K activator 740 Y-P, and the cells were divided into the control group, the 740 Y-P group, the Iso group and the Iso+740 Y-P group. The effects of 740 Y-P on the apoptosis and the expressions of LC3Ⅰ, LC3Ⅱ, p62, PI3K, p-PI3K, mTOR and p-mTOR proteins in each group were detected by flow cytometry and WB. Results: After treatments with 10, 20 and 40 μmol/L Iso, the proliferation capacity of SW480 cells was decreased significantly (all P<0.05); the apoptosis rate was increased significantly (all P<0.05); , the expressions of LC3Ⅱ/LC3Ⅰwere up-regulated significantly (all P<0.05), and the expression of p26 protein was down-regulated significantly (all P<0.05). After treatments with Iso and 740 Y-P, compared with the control group, the apoptosis rate and LC3Ⅱ/LC3Ⅰexpression of the 740 Y-P group were decreased significantly (both P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were increased significantly (all P<0.05).The apoptosis rate and LC3Ⅱ/LC3 expression in the Iso group were increased (both P<0.05) and the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the 740 Y-P group, the apoptosis rate and LC3Ⅱ/LC3Ⅰexpression were increased in the Iso+740 Y-P group (P<0.05), while the expressions of p26, p-PI3K/PI3K and p-mTOR/mTOR were decreased (all P<0.05). Compared with the Iso group, the apoptosis rate and LC3Ⅱ/LC3Ⅰexpression were decreased (both P<0.05), and the expressions of p26, p-PI3K/PI3K, and p-mTOR/mTOR were increased significantly (all P<0.05) in the Iso+740 Y-P group. Conclusion: Iso inhibits the proliferation and induces the apoptosis and autophagy of SW480 cells by inhibiting PI3K/Akt/mTOR signaling pathway.
2024, 31(7):700-706.
DOI: 10.3872/j.issn.1007-385X.2024.07.009
Abstract:
Objective: To explore the effects of homologous recombination repair (HRR) gene mutations on the immunotherapy efficacy and the prognosis of advanced non-small cell lung cancer (NSCLC) patients. Methods: Clinical data of 124 patients with advanced NSCLC who received PD-1 inhibitor treatment between March 2018 and April 2023 at the First Affiliated Hospital of Zhengzhou University were collected. The patients were divided into the mutant group (n=57 cases) and the wild group (n=67 cases) according to the presence or absence of HRR gene mutations. The differences in clinical characteristics and immunotherapy efficacy between the two groups were analyzed by Chi-square test or Fisher's exact test. Kaplan-Meier method was used to compare the progression-free survival (PFS) of the two groups, and univariate and multivariate Cox regression were employed to analyze the factors affecting PFS. Results: The proportions of squamous cell carcinoma and tumor mutation burden (TMB) ≥10 mut/Mb were significantly higher in the HRR gene mutant group than in the wild group (54.4% vs 32.8%, 61.4% vs 29.9%, all PP=0.252), and the disease control rate (DCR) was 86.0% and 73.1% (P=0.080) for patients in the HRR gene mutant group and the wild group, respectively. There was a significant difference in PFS of the HRR gene mutant group and the wild group (6.8 months vs 3.9 months, PHR=0.550, 95%CI [0.352, 0.860], P=0.009) and the number of immunotherapy lines (HR=0.468, 95%CI[0.312, 0.702], PConclusion: The immunotherapy efficacy of the HRR gene mutant group is better than that of the wild group. HRR gene mutations are an independent protective factor for immunotherapy prognosis of patients with advanced NSCLC.
Objective: To explore the effects of homologous recombination repair (HRR) gene mutations on the immunotherapy efficacy and the prognosis of advanced non-small cell lung cancer (NSCLC) patients. Methods: Clinical data of 124 patients with advanced NSCLC who received PD-1 inhibitor treatment between March 2018 and April 2023 at the First Affiliated Hospital of Zhengzhou University were collected. The patients were divided into the mutant group (n=57 cases) and the wild group (n=67 cases) according to the presence or absence of HRR gene mutations. The differences in clinical characteristics and immunotherapy efficacy between the two groups were analyzed by Chi-square test or Fisher's exact test. Kaplan-Meier method was used to compare the progression-free survival (PFS) of the two groups, and univariate and multivariate Cox regression were employed to analyze the factors affecting PFS. Results: The proportions of squamous cell carcinoma and tumor mutation burden (TMB) ≥10 mut/Mb were significantly higher in the HRR gene mutant group than in the wild group (54.4% vs 32.8%, 61.4% vs 29.9%, all PP=0.252), and the disease control rate (DCR) was 86.0% and 73.1% (P=0.080) for patients in the HRR gene mutant group and the wild group, respectively. There was a significant difference in PFS of the HRR gene mutant group and the wild group (6.8 months vs 3.9 months, PHR=0.550, 95%CI [0.352, 0.860], P=0.009) and the number of immunotherapy lines (HR=0.468, 95%CI[0.312, 0.702], PConclusion: The immunotherapy efficacy of the HRR gene mutant group is better than that of the wild group. HRR gene mutations are an independent protective factor for immunotherapy prognosis of patients with advanced NSCLC.
2024, 31(7):707-714.
DOI: 10.3872/j.issn.1007-385X.2024.07.010
Abstract:
Objective: To screen the key genes affecting the prognosis and treatment of gastric cancer (GC) patients and to analyze the value of the key gene microfibrillar-associated protein 2 (MFAP2) in suggesting the prognosis and the sensitivity of immunotherapy of GC patients. Methods: The expression profile data and clinical information of gastric cancer and paracancerous tissues were downloaded from the TCGA database. Genes significantly associated with the prognosis of gastric cancer were screened by weighted gene co-expression network analysis (WGCNA) and univariate Cox regression analysis. Multiple patient cohorts were used to assess the prognostic value of the key gene MFAP2 and analyze its efficacy and clinicopathological correlation. Multiple online databases and algorithms were used to analyze the correlation between MFAP2 and the tumor immune microenvironment. Immunophenotyping score (IPS) combined with immunotherapy patient cohort was used to analyze the value of MFAP2 in predicting the responsiveness of immunotherapy. Differential expression of MFAP2 in gastric cancer and adjacent tissues was verified with multiple datasets. Results: The blue module showed the highest correlation with survival outcomes of GC patients (R=0.17, PHR>1, PPPConclusion: High expression of MFAP2 is a predictive factor for poor prognosis and response to immunotherapy in GC patients. MFAP2 is expected to serve as a novel therapeutic target and prognostic indicator for gastric cancer.
Objective: To screen the key genes affecting the prognosis and treatment of gastric cancer (GC) patients and to analyze the value of the key gene microfibrillar-associated protein 2 (MFAP2) in suggesting the prognosis and the sensitivity of immunotherapy of GC patients. Methods: The expression profile data and clinical information of gastric cancer and paracancerous tissues were downloaded from the TCGA database. Genes significantly associated with the prognosis of gastric cancer were screened by weighted gene co-expression network analysis (WGCNA) and univariate Cox regression analysis. Multiple patient cohorts were used to assess the prognostic value of the key gene MFAP2 and analyze its efficacy and clinicopathological correlation. Multiple online databases and algorithms were used to analyze the correlation between MFAP2 and the tumor immune microenvironment. Immunophenotyping score (IPS) combined with immunotherapy patient cohort was used to analyze the value of MFAP2 in predicting the responsiveness of immunotherapy. Differential expression of MFAP2 in gastric cancer and adjacent tissues was verified with multiple datasets. Results: The blue module showed the highest correlation with survival outcomes of GC patients (R=0.17, PHR>1, PPPConclusion: High expression of MFAP2 is a predictive factor for poor prognosis and response to immunotherapy in GC patients. MFAP2 is expected to serve as a novel therapeutic target and prognostic indicator for gastric cancer.
2024, 31(7):715-721.
DOI: 10.3872/j.issn.1007-385X.2024.07.011
Abstract:
肿瘤免疫逃逸是肿瘤恶性进展的关键阶段，也是肿瘤免疫治疗所面临的一个主要挑战。研究表明，DNA甲基化可通过 影响肿瘤抗原、MHCⅠ类分子、免疫检查点分子和免疫基因的表达，以及巨噬细胞的募集和极化介导肿瘤免疫逃逸的发生与进 展，是肿瘤有前途的治疗靶点。目前，DNA去甲基化药物在肿瘤治疗领域的应用取得了新的进展，包括与免疫检查点抑制剂 （ICI）、其他免疫药物和化疗药物等的联合应用，以及纳米药物和肿瘤疫苗等多种新型药物的开发等。为促进DNA去甲基化药物 作为抗肿瘤药物的发展，本文探讨了DNA甲基化在肿瘤免疫逃逸中的作用机制及DNA去甲基化药物在实验室与临床相关试验 研究现状，提出了改进措施和可能的研究方向。
肿瘤免疫逃逸是肿瘤恶性进展的关键阶段，也是肿瘤免疫治疗所面临的一个主要挑战。研究表明，DNA甲基化可通过 影响肿瘤抗原、MHCⅠ类分子、免疫检查点分子和免疫基因的表达，以及巨噬细胞的募集和极化介导肿瘤免疫逃逸的发生与进 展，是肿瘤有前途的治疗靶点。目前，DNA去甲基化药物在肿瘤治疗领域的应用取得了新的进展，包括与免疫检查点抑制剂 （ICI）、其他免疫药物和化疗药物等的联合应用，以及纳米药物和肿瘤疫苗等多种新型药物的开发等。为促进DNA去甲基化药物 作为抗肿瘤药物的发展，本文探讨了DNA甲基化在肿瘤免疫逃逸中的作用机制及DNA去甲基化药物在实验室与临床相关试验 研究现状，提出了改进措施和可能的研究方向。
2024, 31(7):722-727.
DOI: 10.3872/j.issn.1007-385X.2024.07.012
Abstract:
CAR-T细胞疗法是一种新兴的抗肿瘤免疫细胞疗法，在某些血液系统恶性肿瘤的治疗中取得进展。由于CAR-T细 胞自身存在靶抗原的异质性、自发功能耗竭等问题，同时实体瘤有较强的逃避机体免疫系统的能力，导致单一CAR-T细胞应用于 实体瘤中鲜有成效。CD70是一种Ⅱ型穿膜糖蛋白，在正常人体组织中表达较少或不表达，在多种肿瘤组织中高表达，因此，成为 CAR-T细胞疗法的有效靶点之一。近年来，研究聚焦于靶向CD70的嵌合抗原受体基因修饰T（CD70 CAR-T）细胞在CAR结构 上的不断优化、与其他靶点联合构建双特异性CAR-T细胞（B7H3、CD19等）及联合治疗（PARP抑制剂、溶瘤病毒、表观遗传学调 节剂等），探索增强疗效的新策略。本文论述了CD70 CAR-T细胞疗法的研究进展，分析联合疗法提高疗效的分子机制，为其早 日应用于临床治疗提供了新思路。
CAR-T细胞疗法是一种新兴的抗肿瘤免疫细胞疗法，在某些血液系统恶性肿瘤的治疗中取得进展。由于CAR-T细 胞自身存在靶抗原的异质性、自发功能耗竭等问题，同时实体瘤有较强的逃避机体免疫系统的能力，导致单一CAR-T细胞应用于 实体瘤中鲜有成效。CD70是一种Ⅱ型穿膜糖蛋白，在正常人体组织中表达较少或不表达，在多种肿瘤组织中高表达，因此，成为 CAR-T细胞疗法的有效靶点之一。近年来，研究聚焦于靶向CD70的嵌合抗原受体基因修饰T（CD70 CAR-T）细胞在CAR结构 上的不断优化、与其他靶点联合构建双特异性CAR-T细胞（B7H3、CD19等）及联合治疗（PARP抑制剂、溶瘤病毒、表观遗传学调 节剂等），探索增强疗效的新策略。本文论述了CD70 CAR-T细胞疗法的研究进展，分析联合疗法提高疗效的分子机制，为其早 日应用于临床治疗提供了新思路。
2024, 31(7):728-732.
DOI: 10.3872/j.issn.1007-385X.2024.07.013
Abstract:
曲妥珠单抗是晚期人表皮生长因子受体2（HER2）阳性胃癌患者治疗的一线用药，耐药问题是曲妥珠单抗治疗中面临 的主要挑战。曲妥珠单抗治疗的耐药机制除了与HER2自身状态有关外，也与PI3K/AKT、MEK/ERK等经典信号通路以及有丝 分裂相关的非经典信号通路的异常激活有关，胃癌肿瘤微环境中代谢及免疫调控的改变也会导致患者对曲妥珠单抗耐药，目前 抗体药物偶联物等新型治疗方案可以克服并改善曲妥珠单抗的耐药性。本文聚焦曲妥珠单抗在HER2阳性胃癌中的耐药机制及 其克服曲妥珠单抗耐药新型疗法的研究进展，为临床优化曲妥珠单抗治疗HER2阳性胃癌提供了新思路。
曲妥珠单抗是晚期人表皮生长因子受体2（HER2）阳性胃癌患者治疗的一线用药，耐药问题是曲妥珠单抗治疗中面临 的主要挑战。曲妥珠单抗治疗的耐药机制除了与HER2自身状态有关外，也与PI3K/AKT、MEK/ERK等经典信号通路以及有丝 分裂相关的非经典信号通路的异常激活有关，胃癌肿瘤微环境中代谢及免疫调控的改变也会导致患者对曲妥珠单抗耐药，目前 抗体药物偶联物等新型治疗方案可以克服并改善曲妥珠单抗的耐药性。本文聚焦曲妥珠单抗在HER2阳性胃癌中的耐药机制及 其克服曲妥珠单抗耐药新型疗法的研究进展，为临床优化曲妥珠单抗治疗HER2阳性胃癌提供了新思路。
2024, 31(7):733-737.
DOI: 10.3872/j.issn.1007-385X.2024.07.014
Abstract:
小细胞肺癌（SCLC）是一种预后不良的侵袭性神经内分泌癌，针对SCLC的一线治疗改善有限，目前尚无靶向治疗方 法。Dalta样配体3（DLL3）是Notch信号抑制配体，是一种有吸引力的治疗靶点，其在SCLC细胞表面过表达，而在正常细胞上几 乎不表达。当前正在开发几种DLL3靶向疗法，包括抗体-药物偶联物（ADC）、T细胞衔接器（TCE）分子和嵌合抗原受体基因修饰 （CAR）T细胞疗法，用于治疗SCLC和其他神经内分泌癌。在SCLC治疗中，DLL3靶向ADC药物临床研究结果不理想，但DLL3 靶向TCE和CAR在目前的研究中显示出了有效的抗肿瘤活性。本文回顾了目前正在开发的几种靶向DLL3药物的临床前和临 床试验数据，讨论了DLL3靶向治疗的挑战和机遇，为临床治疗SCLC提供了新思路。
小细胞肺癌（SCLC）是一种预后不良的侵袭性神经内分泌癌，针对SCLC的一线治疗改善有限，目前尚无靶向治疗方 法。Dalta样配体3（DLL3）是Notch信号抑制配体，是一种有吸引力的治疗靶点，其在SCLC细胞表面过表达，而在正常细胞上几 乎不表达。当前正在开发几种DLL3靶向疗法，包括抗体-药物偶联物（ADC）、T细胞衔接器（TCE）分子和嵌合抗原受体基因修饰 （CAR）T细胞疗法，用于治疗SCLC和其他神经内分泌癌。在SCLC治疗中，DLL3靶向ADC药物临床研究结果不理想，但DLL3 靶向TCE和CAR在目前的研究中显示出了有效的抗肿瘤活性。本文回顾了目前正在开发的几种靶向DLL3药物的临床前和临 床试验数据，讨论了DLL3靶向治疗的挑战和机遇，为临床治疗SCLC提供了新思路。
2024, 31(7):738-739.
DOI: 10.3872/j.issn.1007-385X.2024.07.015
Abstract:
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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.