IL-6通过STAT3通路对卵巢癌细胞Bcl-2、CyclinD1和VEGF表达的研究
发布时间：2018-08-22 15:35
【摘要】：目的 讨论白细胞介素-6（Interleukin-6，IL-6）对上皮性卵巢腺癌SKOV3细胞株中的B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、细胞周期蛋白D1（CyclinD1）及血管内皮生长因子（vascular endothelial groeth factor，VEGF）的调控是否与STAT3通路有关。 方法 1、用IL-6浓度为0、10、20、40（ng/ml）分别作用于SKOV3细胞，作用24h、48h后，用MTT法、流式细胞术法检测细胞的增殖和细胞的凋亡；RT-PCR法检测细胞Bcl-2、CyclinD1以及VEGF基因的表达；Western blotting法检测Bcl-2、CyclinD1、VEGF、p-STAT3蛋白的表达。 2、用IL-6浓度为20ng/ml和浓度分别为0、20、40、80、160（μmol/l）的JAK激酶特异性抑制剂AG490联合作用于细胞，用MTT法、流式细胞术法、RT-PCR和Western blotting检测细胞的增殖、凋亡及Bcl-2、CyclinD1、VEGF、p-STAT3的表达。 3、数据用x±s表示，采用SPSS16.0进行数据处理与统计分析，配对样本用t检验，，多样本比较用方差分析。P＜0.05差异有统计学意义。 结果 1、IL-6（0、10、20ng/ml）以浓度依赖的方式，促进细胞的增殖（P＜0.01），抑制其凋亡（P＜0.01），Bcl-2、CyclinD1、VEGF、p-STAT3的蛋白表达量增加（P＜0.01），Bcl-2、CyclinD1、VEGF的基因表达上调（P＜0.01）并于20ng/ml时出现浓度作用峰值。48h和24h差异有统计学意义（P＜0.01）。与IL-6组20ng/ml的比较，40ng/ml组无差异（P＞0.05）。 2、与IL-6组比较，JAK激酶抑制剂AG490组（20、40、80μmol/l）呈剂量依赖性，抑制细胞的增殖（P＜0.01），促进细胞凋亡（P＜0.01），于80μmol/l时作用达峰值。同时，Bcl-2、CyclinD1、VEGF、p-STAT3的基因和蛋白表达明显降低(P＜0.01)。与AG490浓度80μmol/L相比，160μmol/l没有差异性（P＞0.05）。 结论 IL-6可能通过调节STAT3通路，进而调节卵巢癌细胞Bcl-2、CyclinD1、VEGF的转录及表达，从而促进卵巢癌细胞增殖，抑制细胞凋亡。
[Abstract]:Objective to investigate whether the regulation of interleukin-6 (IL-6) on B-cell lymphoma-2 Bcl-2 (BL-2), cyclin D1 (CyclinD1) and vascular endothelial growth factor (vascular endothelial groeth factor-VEGF (VEGF-VEGF-VEGF-VEGF-VEGFR) in SKOV3 cells of epithelial ovarian adenocarcinoma is related to the STAT3 pathway. Methods: (1) SKOV3 cells were treated with IL-6 at the concentration of 0 ~ 10 ~ 20 ~ 40 (ng/ml) for 24 h or 48 h, then the proliferation of cells and the expression of Bcl-2 cyclin D1 and VEGF gene were detected by MTT assay and flow cytometry respectively, and the expression of Bcl-2 cyclin D1 and VEGF gene were detected by RT-PCR. Western blotting assay was used to detect the expression of VEGFP-STAT3 protein in Bcl-2CyclinD1.2The cells were treated with JAK kinase specific inhibitor AG490 with IL-6 concentration of 20ng/ml and JAK kinase specific inhibitor AG490 (渭 mol/l), respectively. The proliferation of the cells was detected by MTT, flow cytometry, RT-PCR and Western blotting. Apoptosis and the expression of VEGF p-STAT3 in Bcl-2 cyclin D1, the data were expressed by x 卤s, the data were processed and statistically analyzed by SPSS16.0, and the paired samples were analyzed by t test. There was significant difference in the comparison of multiple samples with ANOVA (.P < 0. 05). Results 1Interleukin-6 (10ng / ml) was found in a concentration-dependent manner. (P < 0. 01). The protein expression of Bcl-2Cyclin D1 and VEGFU p-STAT3 was increased (P < 0. 01), and the expression of VEGF was up-regulated (P < 0. 01). There was a significant difference between the two groups (P < 0. 01), and the peak value of concentration appeared in 20ng/ml (P < 0. 01) at 48h and 24h (P < 0. 01), and the expression of VEGFP-STAT3 increased significantly (P < 0. 01), and the expression of VEGFP-STAT3 increased significantly (P < 0. 01). There was no difference between 40 ng / ml 20ng/ml and IL-6 group (P > 0. 05). 2. Compared with IL-6 group, AG490 group (20 ~ 4040 ~ 80 渭 mol/l) showed dose-dependent inhibition of cell proliferation (P < 0. 01) and promoted apoptosis (P < 0. 01), and reached its peak at 80 渭 mol/l. At the same time, the gene and protein expression of VEGF p-STAT3 in Bcl-2 cyclin D 1 was significantly decreased (P < 0.01). There was no difference in the concentration of AG490 between 80 渭 mol/L and 160 渭 mol/l (P > 0. 05). Conclusion IL-6 may regulate the transcription and expression of STAT3 in ovarian cancer cells by regulating the STAT3 pathway, thereby promoting the proliferation of ovarian cancer cells and inhibiting apoptosis.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31
[Abstract]:Objective to investigate whether the regulation of interleukin-6 (IL-6) on B-cell lymphoma-2 Bcl-2 (BL-2), cyclin D1 (CyclinD1) and vascular endothelial growth factor (vascular endothelial groeth factor-VEGF (VEGF-VEGF-VEGF-VEGF-VEGFR) in SKOV3 cells of epithelial ovarian adenocarcinoma is related to the STAT3 pathway. Methods: (1) SKOV3 cells were treated with IL-6 at the concentration of 0 ~ 10 ~ 20 ~ 40 (ng/ml) for 24 h or 48 h, then the proliferation of cells and the expression of Bcl-2 cyclin D1 and VEGF gene were detected by MTT assay and flow cytometry respectively, and the expression of Bcl-2 cyclin D1 and VEGF gene were detected by RT-PCR. Western blotting assay was used to detect the expression of VEGFP-STAT3 protein in Bcl-2CyclinD1.2The cells were treated with JAK kinase specific inhibitor AG490 with IL-6 concentration of 20ng/ml and JAK kinase specific inhibitor AG490 (渭 mol/l), respectively. The proliferation of the cells was detected by MTT, flow cytometry, RT-PCR and Western blotting. Apoptosis and the expression of VEGF p-STAT3 in Bcl-2 cyclin D1, the data were expressed by x 卤s, the data were processed and statistically analyzed by SPSS16.0, and the paired samples were analyzed by t test. There was significant difference in the comparison of multiple samples with ANOVA (.P < 0. 05). Results 1Interleukin-6 (10ng / ml) was found in a concentration-dependent manner. (P < 0. 01). The protein expression of Bcl-2Cyclin D1 and VEGFU p-STAT3 was increased (P < 0. 01), and the expression of VEGF was up-regulated (P < 0. 01). There was a significant difference between the two groups (P < 0. 01), and the peak value of concentration appeared in 20ng/ml (P < 0. 01) at 48h and 24h (P < 0. 01), and the expression of VEGFP-STAT3 increased significantly (P < 0. 01), and the expression of VEGFP-STAT3 increased significantly (P < 0. 01). There was no difference between 40 ng / ml 20ng/ml and IL-6 group (P > 0. 05). 2. Compared with IL-6 group, AG490 group (20 ~ 4040 ~ 80 渭 mol/l) showed dose-dependent inhibition of cell proliferation (P < 0. 01) and promoted apoptosis (P < 0. 01), and reached its peak at 80 渭 mol/l. At the same time, the gene and protein expression of VEGF p-STAT3 in Bcl-2 cyclin D 1 was significantly decreased (P < 0.01). There was no difference in the concentration of AG490 between 80 渭 mol/L and 160 渭 mol/l (P > 0. 05). Conclusion IL-6 may regulate the transcription and expression of STAT3 in ovarian cancer cells by regulating the STAT3 pathway, thereby promoting the proliferation of ovarian cancer cells and inhibiting apoptosis.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31
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