腹膜间皮细胞和卵巢癌细胞A2780的相互作用及对顺铂敏感性的变化

Study on the Effect of Human Peritoneal Mesothelial Cells on the Change of the Sensitivity of Ovarian Cancer Cell A2780 to Cisplatin

【目的】探讨卵巢癌细胞A2780与卵巢癌患者的腹膜间皮细胞（HPMC）非接触共培养前后Akt2基因水平改变及对顺铂敏感性的变化。【方法】原代培养卵巢癌患者的HPMC与卵巢癌细胞A2780非接触性共同培养24h，观察细胞形态学变化，收集细胞后，分剐利用半定量Real-timePCR法、Westernblot法测定细胞共培养前后Akt2基因在mR—NA和蛋白水平表达的改变，MTT法检测共培养前后对不同浓度顺铂敏感性的变化。【结果】非接触性共培养后，Akt2mRNA水平表达显著增加（P〈0．05）；总Akt2蛋白培养前后比较无差异（P〉0．05），磷酸化Akt（p-Akt）蛋白明显上调（P〈0．05）；不同浓度顺铂作用后，共培养细胞的存活率升高（P〈O．01）。【结论】HPMC与卵巢癌细胞A2780相互作用可产生促进肿瘤生长的调控因子，引起p-Akt的活化，产生耐药，二者间可能存在着复杂的调控网络。

[Objective] To explore the changes of Akt2 gene expression and sensitivity of ovarian cancer cells A2780 to cisplatin after non-contiguous co-culture of A278 cells and human peritoneal mesothelial cells（HPMC）. [Methods] Primarily cultured HPMC from epithelial ovarian cancers was non-contiguously co-cultured with A2780 cells for 24h. The cell morphol- ogy change was observed. Real-time PCR and western blot were used to detect the Akt2 gene expression changes in mRNA and protein levels in A2780 cells after co-culture. MTT assay was applied to examine the sensitivity changes to cisplatin before and after co-culture. [Results]After non-contiguous co-culture, the Akt2 mRNA expression was increased in A2780 cells. There was no significant difference in total Akt2 expression between before and after co-culture（ P 〉0.05）, but the p-Akt el- evated obviously（ P 〈0.05）. The survival rate of the co-culture cells affected by different concentration of cisplatin was in- creased（ P 〈0.01）. [Conclusion] The interaction between HPMC and ovarian cancer cells A2780 can produce the regulation factor which can promote the tumor group, and activate the p-Akt, and prompt the chemoresistance to cisplatin, so both HPMC and ovarian cancer cells A2780 exist the complex regulation network.