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目的:通过沉默Wnt/β-catenin信号转导通路的核心蛋白β-catenin,探讨顺铂对人卵巢癌SKOV3细胞增殖、凋亡的影响,为卵巢癌的耐药逆转提供新思路。方法:用包含β-catenin基因特异RNA序列的慢病毒转染人卵巢癌SKOV3细胞,沉默β-catenin蛋白表达。分组:SKOV3组、SKOV3-neg组(转染空载体)、SKOV3-RNAi组(转染β-catenin-RNAi慢病毒)。Western blot法检测转染前后SKOV3细胞中β-catenin蛋白的表达。12.5μg/ml顺铂作用48h后,用MTT法检测顺铂对各组SKOV3细胞增殖抑制率的影响;流式细胞术检测各组中SKOV3细胞的凋亡率。结果:(1)Western blot结果显示,SKOV3-RNAi组中β-catenin蛋白表达水平显著低于SKOV3-neg和SKOV3组(P<0.05)。(2)相同浓度顺铂作用下,SKOV3-RNAi组的增殖抑制率显著高于SKOV3组及SKOV3-neg组(P<0.05),而后两组的增殖抑制率无显著差异(P>0.05)。(3)流式细胞术检测结果显示,SKOV3-RNAi、SKOV3-neg和SKOV3组的细胞凋亡率分别为(18.8±4.3)%、(1.8±0.3)%、(1.9±0.3)%,SKOV3-RNAi组凋亡率显著高于其余两组(P=0.008<0.05)。顺铂作用后,SKOV3-RNAi、SKOV3-neg和SKOV3组的细胞凋亡率分别为(67.7±2.5)%、(33.3±2.6)%、(32.3±2.1)%;顺铂作用后,各组凋亡率均显著升高(P均<0.05),且SK-OV3-RNAi组凋亡率显著高于其余两组(P=0.000<0.05)。结论:沉默β-catenin蛋白能有效增强SKOV3对cDDP的敏感性,β-catenin有可能成为逆转卵巢癌化疗耐药的治疗靶点。
OBJECTIVE: To investigate the effect of cisplatin on the proliferation and apoptosis of human ovarian cancer cell line SKOV3 by silencing the core protein β-catenin of Wnt / β-catenin signaling pathway, and to provide new ideas for the reversal of drug resistance in ovarian cancer. Methods: Human ovarian cancer SKOV3 cells were transfected with lentivirus containing the specific RNA of β-catenin gene to silence the expression of β-catenin protein. Grouping: SKOV3 group, SKOV3-neg group (empty vector transfected), SKOV3-RNAi group (transfected with β-catenin-RNAi lentivirus). Western blot was used to detect the expression of β-catenin protein in SKOV3 cells before and after transfection. After treated with 12.5μg / ml cisplatin for 48h, the effect of cisplatin on the proliferation inhibition rate of SKOV3 cells was detected by MTT assay. The apoptosis rate of SKOV3 cells in each group was detected by flow cytometry. Results: (1) Western blot results showed that the expression ofβ-catenin in SKOV3-RNAi group was significantly lower than that in SKOV3-neg and SKOV3 groups (P <0.05). (2) Under the same concentration of cisplatin, the proliferation inhibition rate in SKOV3-RNAi group was significantly higher than that in SKOV3 and SKOV3-neg groups (P <0.05), but no significant difference was found between the two groups (P> 0.05). (3) The results of flow cytometry showed that the apoptosis rates of SKOV3-RNAi, SKOV3-neg and SKOV3 groups were (18.8 ± 4.3)%, (1.8 ± 0.3)%, (1.9 ± 0.3)% and -RNAi group apoptosis rate was significantly higher than the other two groups (P = 0.008 <0.05). The apoptosis rates of SKOV3-RNAi, SKOV3-neg and SKOV3 group were (67.7 ± 2.5)%, (33.3 ± 2.6)% and (32.3 ± 2.1)% respectively after cisplatin treatment. (P <0.05), and the apoptosis rate in SK-OV3-RNAi group was significantly higher than the other two groups (P = 0.000 <0.05). CONCLUSION: Silencing β-catenin protein can effectively enhance the sensitivity of SKOV3 to cDDP. Β-catenin may be a therapeutic target for reversing the chemoresistance of ovarian cancer.