论文部分内容阅读
目的观察MEK1/2特异性抑制剂PD98059联合顺铂对卵巢癌SKOV3细胞增殖的影响。方法按不同处理因素对卵巢癌SKOV3细胞分组:空白对照组、单纯PD98059组、单纯顺铂组、联合用药组。CCK-8法检测各组细胞的增殖抑制率。实时定量PCR检测各组细胞含有WW结构域的氧化还原酶(WWOX)和细胞外调节蛋白激酶(ERK)在基因水平的表达。Western-blot检测各组细胞磷酸化ERK1/2(p-ERK1/2)蛋白的表达。光镜下观察各组细胞的生长状态及形态学改变。流式细胞术检测各组细胞的凋亡情况。结果联合用药组细胞增殖抑制率明显高于其他各组(P<0.05);单纯顺铂组及联合用药组WWOX mRNA表达量较空白对照组明显增加(P<0.05),单纯PD98059组WWOX表达量无明显改变(P>0.05);ERK mRNA在各组间的表达无明显变化(P>0.05);与空白对照组相比,p-ERK1/2表达量在单纯PD98059组及联合用药组中明显降低(P<0.05),在单纯顺铂组的表达无明显变化。光镜下可观察到空白对照组及单纯PD98059组细胞生长状态好,细胞密集,无明显形态学改变;单纯顺铂组及联合用药组贴壁细胞少,细胞稀疏,联合用药组部分细胞出现胞浆溶解样改变。结论 PD98059联合顺铂可能通过调控WWOX基因表达及ERK通路,对卵巢癌细胞增殖产生较强的抑制作用。
Objective To observe the effect of MEK1 / 2 specific inhibitor PD98059 and cisplatin on the proliferation of ovarian cancer SKOV3 cells. Methods SKOV3 cells were divided into different groups according to different treatment factors: blank control group, PD98059 group, cisplatin alone group and combination group. CCK-8 method was used to detect the proliferation inhibition rate of each group of cells. Real-time quantitative PCR was used to detect the WWOX and ERK expression in WW cells. The expression of phosphorylated ERK1 / 2 (p-ERK1 / 2) protein in each group was detected by Western-blot. Under light microscope, the growth and morphological changes of each group were observed. Flow cytometry was used to detect the apoptosis of each group of cells. Results Compared with other groups, the inhibition rate of WWOX in combination group was significantly higher than that in other groups (P <0.05). The WWOX mRNA expression in combination cisplatin group and combination group was significantly higher than that in blank control group (P <0.05). WWOX expression in PD98059 group (P> 0.05). The expression of ERK mRNA did not change significantly between the groups (P> 0.05). Compared with the blank control group, the expression of p-ERK1 / 2 was significantly increased in the PD98059 group and the combination group (P <0.05), but there was no significant difference in cisplatin alone. Under the light microscope, cells in blank control group and PD98059 group showed good cell growth and dense cells with no obvious morphological changes. Adherent cells in the cisplatin group and the combination group had few adherent cells and sparse cells. Some cells in the combination group showed cells Slurry lysis changed. Conclusion PD98059 combined with cisplatin may inhibit the proliferation of ovarian cancer cells through regulating WWOX gene expression and ERK pathway.