PEITC对NSCLC A549、H1299细胞凋亡的影响

The effect of PEITC on apoptosis of NSCLC A549 and H1299 cells

目的 探讨异硫氰酸苯乙酯(PEITC)对人非小细胞肺癌(NSCLC)细胞株(A549、H1299)凋亡的影响机制。方法 体外培养A549、H1299细胞,采用10、30、50μmol/L PEITC处理细胞48 h后,MTT法检测细胞增殖水平,流式细胞术检测细胞周期及凋亡率。实时定量PCR、Western blotting分别检测细胞内TFPI-2 mRNA及蛋白、TFPI-2甲基化、TMPRSS4表达水平。荧光素酶报告实验检测细胞内TMPRSS4活性。沉默TFPI-2或过表达TMPRSS4后,分别检测肺癌细胞增殖率及凋亡率。结果 不同浓度PEITC能够抑制A549、H1299细胞增殖及诱导细胞凋亡。PEITC能够促进细胞TFPI-2 mRNA以及蛋白的表达,降低TFPI-2的甲基化酶活性及TMPRSS4的表达。沉默TFPI-2表达后,A549、H1299细胞增殖水平显著增加,而过表达TMPRSS4降低细胞凋亡率。结论 PEITC能够诱导NSCLC A549、H1299细胞凋亡,其机制可能与抑制TFPI-2甲基化、下调TMPRSS4表达有关。

Aim To investigate the mechanism of the effect of phenethyl isothiocyanate(PEITC) on apoptosis in human non-small cell lung cancer(NSCLC) cell lines(A549,H1299).Methods A549 and H1299 cells were cultured in vitro and treated with 10、30、50 μmol/L of PEITC for 48 h,cell proliferation was measured by MTT assay and flow cytometry were used to detect cell cycle and apoptosis rate.The expression of TFPI-2 mRNA and protein,TFPI-2 methylation and TMPRSS4 were detected by real-time quantitative PCR and Western blotting.Fluorescence reporter gene assay was used to detect the activity of TMPRSS4 reporter gene in cells.After silencing TFPI-2 or overexpressing TMPRSS4,the proliferation rate and apoptosis rate of lung cancer cells were detected.Results Different concentrations of PEITC can inhibit the proliferation and induce apoptosis of A549 and H1299 cells.In addition,PEITC can also promote the expression of TFPI-2 mRNA and protein in cells,and the activity of TFPI-2 methylase and the expression of TMPRSS4 in cells were decreased.Silencing the expression of TFPI-2 significantly increased the proliferation level of A549 and H1299 cells,while overexpression of TMPRSS4 decreased the apoptosis rate of cells.Conclusion PEITC can induce apoptosis in NSCLC A549 and H1299 lung cancer cells,which may be related to inhibition of TFPI-2 methylation and downregulation of TMPRSS4 expression.