千金藤素诱导人肺腺癌LTEP-a-2细胞中miRNA表达变化的研究

Cepharanthine induces miRNA expression in human lung adenocarcinoma LTEP-a-2 cells

目的:观察千金藤素对人肺腺癌LTEP-a-2细胞生长的影响,探讨细胞中相关微小RNA(miRNA)的表达变化,为明确千金藤素的抗肿瘤机制提供科学依据。方法:在0μmol/L、10μmol/L、20μmol/L和40μmol/L的千金藤素作用后,MTT法检测LTEP-a-2细胞的生长抑制率;光镜下观察千金藤素作用后LTEP-a-2细胞的形态学变化;流式细胞术分析不同浓度千金藤素对LTEP-a-2细胞凋亡的影响;提取细胞miRNA,real-time PCR检测不同浓度千金藤素作用之后细胞中的let-7c、miR-34a和miR-34b的表达情况。结果:MTT法检测结果表明,千金藤素能够抑制LTEP-a-2细胞活力,呈现剂量依赖性;倒置显微镜下可见,随着千金藤素浓度的升高,细胞固缩的程度逐渐加深;流式细胞术分析发现,不同浓度千金藤素均可导致的LTEP-a-2细胞凋亡率增加;real-time PCR结果表明,千金藤素可以使LTEP-a-2细胞中的let-7c、miR-34a和miR-34b表达升高。结论:千金藤素可以抑制LTEP-a-2细胞生长,诱导细胞凋亡;千金藤素升高细胞中let-7c、miR-34a和miR-34b表达,提示这些miRNA在LTEP-a-2细胞具有抑癌基因的功能。

AIM： To observe the effect of cepharanthine on human lung adenocarcinoma LTEP-a-2 cell growth,and to explore the changes of related microRNA（ miRNA） expression in the cells. METHODS： LTEP-a-2 cells were treated with cepharanthine at concentrations of 0 μmol / L,10 μmol / L,20 μmol / L and 40 μmol / L. The growth inhibition rate was detected by MTT assay,and the cell morphological changes were observed under light microscope. The cell apoptosis was analyzed by flow cytometry. The expression of let-7c,miR-34 a and miR-34 b was measured by real-time PCR. RESULTS： Cepharanthine inhibited the cell activity of LTEP-a-2 cells in a dose-dependent manner. With the increase in cepharanthine concentration,the pyknosis of the cells was visible under the inverted microscope. Flow cytometry analysis found that different concentrations of cepharanthine induced the increase in the apoptotic rates of LTEP-a-2 cells. The results of real-time PCR showed that the cepharanthine also increased the expression of let-7c,miR-34 a and miR-34 b. CONCLUSION： Cepharanthine inhibits the growth of LTEP-a-2 cells,and induces apoptosis. Cepharanthine increases the expression of let-7c,miR-34 a and miR-34 b,indicating that these miRNAs in LTEP-a-2 cells has the function as tumor suppressor genes.