罗米地辛与阿糖胞苷联合用药在肺腺癌细胞中介导组蛋白H3K14乙酰化提高CASP3转录活性

Combination of Romidepsin and Cytarabine Enhances CASP3 Transcriptional Activity Through H3K14 Acetylation in Lung Adenocarcinoma Cells

肺癌发病率和病死率均居全球恶性肿瘤首位,有效的药物是治疗肺癌的关键。罗米地辛和阿糖胞苷是已经批准上市并应用于临床治疗的癌症化疗药物。这两种药物的联合用药是否在肺腺癌的治疗中发挥作用及其相关分子机制尚不明确。本研究通过MTT和克隆形成实验证实,罗米地辛和阿糖胞苷联合用药可以显著抑制肺腺癌细胞A549的生长(P<0.05)。体外Transwell细胞侵袭实验和划痕实验表明,联合用药可有效降低肺腺癌细胞的侵袭和转移能力(P<0.05)。同时,流式细胞分析显示,联合用药可以显著诱导肺腺癌细胞的凋亡(P<0.05),并且实时PCR和Western印迹结果提示,肺腺癌细胞A549在联合用药处理后,凋亡相关蛋白质CASP3的mRNA和蛋白质表达水平均明显升高。另外,通过检测联合用药后,A549细胞的组蛋白乙酰化修饰情况发现,H3K14位点乙酰化水平被明显上调,并且ChIP分析进一步显示,CASP3启动子区域组蛋白H3K14乙酰化水平在联合用药后明显增加(P<0.05)。本研究揭示了罗米地辛与阿糖胞苷联合用药能通过增强CASP3启动子区组蛋白H3K14乙酰化水平进而促进CASP3的转录和蛋白质表达,从而抑制肺腺癌细胞的生长、增殖、迁移和侵袭,促进其凋亡的分子机制。在分子和细胞水平上为罗米地辛和阿糖胞苷联合用药治疗肺腺癌提供依据,并为肺腺癌的临床治疗提供切实参考。

Lung cancer ranks first for both incidence and mortality among all types of cancer worldwide.Effective drugs are one of the key to lung cancer treatment.Romidepsin and cytarabine have been approved for clinical treatment as cancer chemotherapeutic drugs.It is not clear yet whether the combination of these two drugs plays a role in the treatment of lung adenocarcinoma and how the molecular mechanism of the combination of romidepsin and cytarabine is in cancer treatment.In this study,we found that the combination of romidepsin and cytarabine induced growth arrest of lung adenocarcinoma A549 cells by MTT and clone formation assays(P<0.05).Also,the invasiveness and metastasis ability of A549 cells were effectively reduced after stimulation with the combination of romidepsin and cytarabine.In the meanwhile,the combination treatment induced apoptosis of A549 cells was detected by flow cytometry assay.In addition,the combination of romidepsin and cytarabine increased the expression of CASP3 in both mRNA and protein levels(P<0.05).Furthermore,we found that the total level of H3K14 acetylation was significantly increased and the acetylation of H3K14 was recruited onto the promoter of CASP3 after the combination treatment.Together,our data reveal a novel mechanism by which the combination of romidepsin and cytarabine activates CASP3 expression through regulating H3K14 acetylation near the CASP3 promoter to inhibit the growth of A549 cells.This research will provide information for finding potential treatment for lung adenocarcinoma.