Decorin基因表达对人肺腺癌A549细胞生物学行为的影响

The influences of Decorin gene on biological behavior in human lung adenocarcinoma A549 cell line

目的探讨Decorin表达降低对人肺腺癌A549细胞的生物学行为的影响。方法体外化学合成DCN-siRNA特异性序列,在Lipofectamine TM2000的介导下转染人肺腺癌A549细胞;分别采用RT-PCR法和Western blot方法检测Decorin基因和蛋白表达情况;CCK-8法检测细胞的增殖能力;流式细胞技术检测细胞凋亡;Transwell小室测定细胞侵袭能力;划痕实验检测肿瘤细胞迁移能力。结果 RT-PCR及Western blot证实靶向Decorin的siRNA干扰明显降低了A549细胞Decorin mRNA和蛋白表达水平。CCK-8检测结果显示与对照组比较,siRNA干扰组A549细胞增殖能力增强,差异具有统计学意义(P<0.05)。流式细胞技术显示,DCN-siRNA组与对照组比较凋亡率差异无统计学意义(P=0.214)。Transwell侵袭实验显示siRNA干扰后,A549细胞穿膜细胞数明显多于对照组(P<0.05)。细胞划痕实验显示与空载体组比较,DCN-siRNA组细胞48 h、72 h细胞生长明显增多,愈合加快。结论应用siRNA技术在肺腺癌细胞中下调Decorin表达促进了A549细胞的增殖,迁移以及侵袭能力,不影响细胞的凋亡能力。该结果为Decorin在肺癌的作用及功能方面的研究提供了新的证据。

Objective To explore the influences of decorin （ DCN） gene down-regulating on biological behavior in human lung adenocarcinoma A 549 cell line.Methods Chemically synthesis in vitro targeting DCN -siRNA was transfected into human lung adenocarcinoma A 549 cells by LipofectamineTM 2000 .The gene expres-sion of DCN was detected using Real-Time PCR ;The protein expression of DCN was investigated using Western blot;Checkout DCN-siRNA effects on lung adenocarcinoma cancer cell proliferation was detected by CCK -8;The migration and invasion ability were determined by Transwell assay .Results DCN-siRNA was successfully transfected into human lung adenocarcinoma A 549 cells.Real-Time PCR results showed that DCN mRNA ex-pression level significantly decreased ,compared with untransfected group ,negative control group .Western blot re-sults showed that DCN-siRNA transfection inhibited DCN protein expression level;CCK-8 results showed that the proliferation was enhanced in the DCN -siRNA group as compared with untransfected group ,as well as the negative control group and empty vector group .Transwell assay results showed the invasion of A 549 cells in DCN-siRNA group was significantly enhanced as compared with untransfected group [（22.6 ±1.14） vs.（5.2 ± 0.84）].Wound-Healing assay results revealed that the cell repairing rate was markedly increased in DCN -siRNA group.A549 cells were close to complete repair after 48 hours.Apoptosis was not significant in DCN -siR-NA group as compared with untransfected group （P=0.214）.Conclusion Down-regulation the expression of DCN gene by DCN-siRNA may enhance the ability of invasion ,migration and proliferation of A 549 cells without＆nbsp;affecting apoptosis ,which provides a new evidence of function for advancing research of lung cancer .