摘要
【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.
出处
《医学临床研究》
CAS
2011年第4期596-599,共4页
Journal of Clinical Research
基金
[基金项目]国家自然科学基金(No.30973184)和国家“973”计划(No.2009CB521800)
关键词
卵巢肿瘤
间质细胞
腹膜
顺铂
ovarian neoplasms
stromal cells
peritoneum
cisplatin