灵芝孢子油对人肺腺癌LTEP-a2细胞凋亡的影响

Glossy Ganoderma Spore Oil Induced Apoptosis of Human Lung Adenocarcinoma LTEP-a2 Cells through Up-regulating miR-16 Expression

目的观察灵芝孢子油对人肺腺癌细胞LTEP-a2增殖、凋亡与miR-16及其靶基因表达的影响,从miRNA角度揭示灵芝孢子油的抗肿瘤机制。方法灵芝孢子油处理人肺腺癌细胞LTEP-a2 24和48 h后,采用CCK-8法检测其对细胞增殖的影响,倒置相差显微镜观察细胞形态学变化,流式细胞术检测细胞凋亡情况,real-time PCR检测miR-16及其靶基因bcl2和血管内皮生长因子(VEGF)的表达。结果灵芝孢子油呈剂量、时间性地抑制人肺腺癌LTEP-a2细胞的增殖;当灵芝孢子油浓度达到2μL.mL-1,细胞形态发生明显变化;Annexin-V/PI双染检测提示,灵芝孢子油在低浓度下就可促进细胞凋亡;灵芝孢子油可显著上调LTEP-a2细胞中miR-16的表达并相应下调bcl2和VEGF基因的表达。结论灵芝孢子油能显著抑制人肺腺癌细胞LTEP-a2细胞增殖,引起细胞形态学改变。其作用机制之一是上调miR-16的表达、下调Bcl2和VEGF的表达,从而促进肺癌细胞凋亡,达到抗肿瘤的效果。

Objective To investigate the effects of glossy gnoderma sore oil on the proliferation,apoptosis,miR-16 expression of human lung adenocarcinoma LTEP-a2 cell line,and explore its anti-cancer mechanism. Methods The LTEP-a2 cells were treated with glossy ganoderma spore oil for 24 and 48 hours.The inhibition on cell growth was determined by using cell count kit（CCK-8）,cell morphological changes were observed by light microscopy,cell apoptosis was analyzed by flow cytometry,and the expression of miR-16,Bcl2 and PDCD4 were determined by real-time PCR. Results Glossy ganoderma spore oil time-and dose-dependently inhibited the LTEP-a2 cells proliferation;changed the cell morphology obviously after the glossy ganoderma spore oil reached to 2 μL·mL-1.Glossy ganoderma spore oil induced the LTEP-a2 cells apoptosis even at a low concentration by Annexin-V/PI double stained detection;and it up-regulated the expression of miR-16 and down-regulated the bcl2 and VEGF expression significantly. Conclusion Glossy ganoderma spore oil could inhibit the cell proliferation obviously and change the cell morphology,the mechanisms of which should be associated with up-regulating the miR-16 expression and down-regulating the bcl2 and VEGF expression.