XIAP gene downregulation by small interfering RNA inhibits proliferation, induces apoptosis, and reverses the cisplatin resistance of ovarian carcinoma
XIAP gene downregulation by small interfering RNA inhibits proliferation, induces apoptosis, and reverses the cisplatin resistance of ovarian carcinoma
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DOI：
10.1016/j.ejogrb.2009.06.011
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发表时间：
2009-10-01
影响因子：
2.6
通讯作者：
Xin, Xiao-yan
中科院分区：
文献类型：
--
作者：
Ma, Jia-jia;Chen, Bi-liang;Xin, Xiao-yan
目的：XIAP是凋亡抑制剂家族最重要的成员之一。它在包括人类卵巢癌在内的多种恶性肿瘤中表达上调，促进恶性细胞的侵袭、转移、生长和存活，并且还赋予对某些化疗药物的耐药性。我们观察了XIAP基因RNA干扰（RNAi）对人卵巢癌细胞系A2780/cp70的增殖、凋亡、致瘤性和化疗敏感性的影响。研究设计：我们使用人U6启动子驱动的DNA模板方法诱导短发夹RNA触发的RNAi来阻断人卵巢癌细胞系A2780/cp70中XIAP基因的表达。构建相应的位点突变载体作为阴性对照（pSilencer 2.1-NC）。 RT-PCR和Western blotting检测稳转细胞和未转染细胞中XIAP mRNA和蛋白的表达。检查细胞生长并使用MTT测定法测定药物敏感性。流式细胞术检测细胞凋亡，裸鼠致瘤性检测。结果：建立了3个稳定转染细胞系：A2780/cp70-s2、A2780/cp70-NC、A2780/cp70-neo。 A2780/cp70-s2中XIAP基因mRNA和蛋白的表达水平分别为63.3%和60.8%，低于A2780/cp70-NC、A2780/cp70-neo和未转染的A.2780/cp70细胞。 A2780/cp70-s2 的细胞增殖减少高达 62.2 +/- 1.9%。与其他细胞系相比，A2780/cp70-s2细胞凋亡率变化为31.1±1.3%，明显增加（P < 0.05）。 XIAP 的敲低可延缓裸鼠的致瘤性，21 天后，A2780/cp70-s2 组的平均肿瘤重量降低了 1.62 +/- 0.11 g（P < 0.05）。添加1μM和10μM顺铂后，A2780/cp70-s2细胞的存活指数明显降低（P < 0.05）。结论：XIAP基因RNIAi通过降低其mRNA和蛋白，抑制卵巢癌细胞的增殖，使细胞对顺铂更加敏感。这些结果表明XIAP是卵巢癌相关基因并且XIAP是治疗性抗癌药物的潜在靶点。 (C) 2009 Elsevier Ireland Ltd. 保留所有权利。
Objective: XIAP is one of the most important members of the inhibitors of apoptosis family. It is upregulated in various malignancies, including human ovarian carcinomas, it promotes invasion, metastasis, growth, and survival of malignant cells, and it also confers resistance to some chemotherapeutic drugs. We observed the effect of XIAP gene RNA interference (RNAi) on the proliferation, apoptosis, tumorigenicity, and chemosensitivity of the human ovarian carcinoma cells A2780/cp70.Study design: We used a human U6 promoter-driven DNA template approach to induce short hairpin RNA-triggered RNAi to block XIAP gene expression in the human ovarian cancer cell line A2780/cp70. A corresponding site-mutated vector was constructed as a negative control (pSilencer 2.1-NC). The expression of XIAP mRNA and protein among the stable transfected cells and the untransfected ones was detected by RT-PCR and Western blotting. The cell growth was examined and drug sensitivity was assayed using the MTT assay. Cell apoptosis was measured by flow cytometry and the tumorigenicity by using nude mice.Results: Three stable transfected cell lines: A2780/cp70-s2, A2780/cp70-NC, and A2780/cp70-neo were established. The expression levels of XIAP gene mRNA and protein in A2780/cp70-s2 were 63.3% and 60.8%, lower than in A2780/cp70-NC, A2780/cp70-neo, and untransfected A.2780/cp70 cells. The cell proliferation of A2780/cp70-s2 was reduced by up to 62.2 +/- 1.9%. Compared with the other cell lines, the changes in apoptotic rate in A2780/cp70-s2 was 31.1 +/- 1.3%, obviously increasing (P < 0.05). The knockdown of XIAP retards tumorigenicity in nude mice and after 21 days the average tumor weight of the A2780/cp70-s2 group was lower by 1.62 +/- 0.11 g (P < 0.05). And the survival index in A2780/cp70-s2 cells was markedly reduced with the addition of 1 mu M and 10 mu M cisplatinum, respectively (P < 0.05).Conclusions: XIAP gene RNIAi inhibited the proliferation of ovarian carcinoma cells and caused cells to be more sensitive to cisplatin through the reduction of its mRNA and protein. These results suggest that XIAP is an ovarian cancer-related gene and that XIAP is a potential target for therapeutic anti-cancer drugs. (C) 2009 Elsevier Ireland Ltd. All rights reserved.