【摘要】：目的 討論白細(xì)胞介素-6（Interleukin-6，IL-6）對(duì)上皮性卵巢腺癌SKOV3細(xì)胞株中的B淋巴細(xì)胞瘤-2（B-cell lymphoma-2，Bcl-2）、細(xì)胞周期蛋白D1（CyclinD1）及血管內(nèi)皮生長(zhǎng)因子（vascular endothelial groeth factor，VEGF）的調(diào)控是否與STAT3通路有關(guān)。 方法 1、用IL-6濃度為0、10、20、40（ng/ml）分別作用于SKOV3細(xì)胞，作用24h、48h后，用MTT法、流式細(xì)胞術(shù)法檢測(cè)細(xì)胞的增殖和細(xì)胞的凋亡；RT-PCR法檢測(cè)細(xì)胞Bcl-2、CyclinD1以及VEGF基因的表達(dá)；Western blotting法檢測(cè)Bcl-2、CyclinD1、VEGF、p-STAT3蛋白的表達(dá)。 2、用IL-6濃度為20ng/ml和濃度分別為0、20、40、80、160（μmol/l）的JAK激酶特異性抑制劑AG490聯(lián)合作用于細(xì)胞，用MTT法、流式細(xì)胞術(shù)法、RT-PCR和Western blotting檢測(cè)細(xì)胞的增殖、凋亡及Bcl-2、CyclinD1、VEGF、p-STAT3的表達(dá)。 3、數(shù)據(jù)用x±s表示，采用SPSS16.0進(jìn)行數(shù)據(jù)處理與統(tǒng)計(jì)分析，配對(duì)樣本用t檢驗(yàn)，，多樣本比較用方差分析。P＜0.05差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果 1、IL-6（0、10、20ng/ml）以濃度依賴(lài)的方式，促進(jìn)細(xì)胞的增殖（P＜0.01），抑制其凋亡（P＜0.01），Bcl-2、CyclinD1、VEGF、p-STAT3的蛋白表達(dá)量增加（P＜0.01），Bcl-2、CyclinD1、VEGF的基因表達(dá)上調(diào)（P＜0.01）并于20ng/ml時(shí)出現(xiàn)濃度作用峰值。48h和24h差異有統(tǒng)計(jì)學(xué)意義（P＜0.01）。與IL-6組20ng/ml的比較，40ng/ml組無(wú)差異（P＞0.05）。 2、與IL-6組比較，JAK激酶抑制劑AG490組（20、40、80μmol/l）呈劑量依賴(lài)性，抑制細(xì)胞的增殖（P＜0.01），促進(jìn)細(xì)胞凋亡（P＜0.01），于80μmol/l時(shí)作用達(dá)峰值。同時(shí)，Bcl-2、CyclinD1、VEGF、p-STAT3的基因和蛋白表達(dá)明顯降低(P＜0.01)。與AG490濃度80μmol/L相比，160μmol/l沒(méi)有差異性（P＞0.05）。 結(jié)論 IL-6可能通過(guò)調(diào)節(jié)STAT3通路，進(jìn)而調(diào)節(jié)卵巢癌細(xì)胞Bcl-2、CyclinD1、VEGF的轉(zhuǎn)錄及表達(dá)，從而促進(jìn)卵巢癌細(xì)胞增殖，抑制細(xì)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 鹵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.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R737.31

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