目的 探究靶向沉默野生型p53诱导的磷酸酶1(Wip1)对卵巢癌细胞紫杉醇(PTX)化疗敏感性的影响.方法 以卵巢癌A2780细胞为研究对象,通过慢病毒感染A2780细胞,构建Wip1低表达的稳定细胞株 Wip1-sgRNA(Wip1-sgRNA组),对照细胞株为NC-sgRNA(NC-sgRNA组),并适时以PTX处理细胞(设为Wip1-sgRNA+PTX组和NC-sgRNA+PTX组);采用CCK8法检测细胞增殖能力,流式细胞术检测细胞凋亡情况,Transwell侵袭实验检测细胞侵袭能力.结果 与NC-sgRNA组比较,Wip1-sgRNA组细胞 Wip1 mR-NA及蛋白表达水平明显下调(P＜0.001);在PTX梯度处理相同时间的条件下,Wip1-sgRNA组细胞存活率均明显低于NC-sgRNA组(P＜0.05);PTX对 Wip1-sgRNA组细胞的48 h时半抑制浓度(IC50)明显低于NC-sgRNA 组(P＜0.001).与NC-sgRNA组比较,Wip1-sgRNA组细胞增殖能力明显减弱(P＜0.001),细胞凋亡率明显升高(P＜0.001),穿膜细胞数明显减少(P＜0.001);PTX处理后,细胞增殖能力均较各自对照组减弱,细胞凋亡率均升高,穿膜细胞数均减少,并且与NC-sgRNA+PTX组比较,Wip1-sgRNA+PTX组细胞增殖能力明显减弱(P＜0.001),细胞凋亡率明显升高(P＜0.001),穿膜细胞数明显减少(P＜0.001).结论 Wip1基因沉默可能提高人卵巢癌A2780细胞对PTX的化疗敏感性.
Objective To explore the effect of targeted silencing wild-type p53-induced phosphatase 1(Wip1)on paclitaxel(PTX)chemosensitivity of ovarian cancer cell line A2780.Methods The ovarian cancer cell lines A2780 served as the research object,the stable cell line Wip1-sgRNA with low expression of Wip1 was constructed by infecting cell line A2780 with lentivirus(Wip1-sgRNA group),while the control cell line was NC-sgRNA(NC-sgRNA group).The cells were treated with PTX at appropriate time and divided into the Wip1-sgRNA+PTX group and NC-sgRNA+PTX group.The CCK8 method was used to detect the cell proliferation ability,the flow cytometry was used to detect the cell apoptosis,and the Transwell invasion assay was used to detect the cell invasion ability.Results Compared with the NC-sgRNA group,the expression lev-els of Wip1 mRNA and protein in the Wip1-sgRNA group were significantly down-regulated(P＜0.001).Un-der the condition of PTX gradient treatment for the same time,the cell survival rate of the Wip1-sgRNA group was significantly lower than that of the NC-sgRNA group(P＜0.001).The half maximal inhibitory concentration(IC50)of PTX in cells of the Wip1-sgRNA group at 48 h was significantly lower than that of the NC-sgRNA group(P＜0.001).Compared with the NC-sgRNA group,the proliferation ability of the Wip1-sgRNA group was significantly weakened(P＜0.001),the rate of cellular apoptosis was significantly in-creased(P＜0.001),and the number of transmenbrane cells was significantly decreased(P＜0.001).After PTX treatment,the proliferation ability was weakened compared with corresponding control groups,the rate of cellular apoptosis was significantly increased,and the number of transmenbrane cells was significantly de-creased;additionally,compared with the NC-sgRNA+PTX group,the cellular proliferation ability of the Wip1-sgRNA+PTX group was significantly weakened(P＜0.001),the rate of cellular apoptosis was signifi-cantly increased(P＜0.001),and the number of transmenbrane cells was significantly decreased(P＜0.001).Conclusion Wip1 gene silencing could increase the chemosensitivity of human ovarian cancer cell lines A2780 to PTX.
裴乃枢;李宗涛;顾笑梅;周麟丰
唐山市妇幼保健院妇产科,河北唐山 063000||华北理工大学,河北唐山 063210||唐山市发育异常与相关疾病分子医学重点实验室,河北唐山 063210华北理工大学附属医院乳腺外科,河北唐山 063000唐山市妇幼保健院妇产科,河北唐山 063000华北理工大学,河北唐山 063210||唐山市发育异常与相关疾病分子医学重点实验室,河北唐山 063210||唐山市妇幼保健院检验科,河北唐山 063000
临床医学
卵巢癌;野生型p53诱导的磷酸酶1;紫杉醇;化疗敏感性;细胞增殖;细胞凋亡;侵袭
ovarian neoplasms;wild-type p53-induced phosphatase 1;paclitaxel;chemosensitivity;pro-liferation;apoptosis;invasion
《重庆医学》 2024 (013)
1921-1926 / 6
河北省自然科学基金项目(H2021105005).
10.3969/j.issn.1671-8348.2024.13.001
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