Decorin基因表达对人肺腺癌A549细胞生物学行为的影响

doi:10.11904/j.issn.1002-3070.2016.04.001

实用肿瘤学杂志 ›› 2016, Vol. 30 ›› Issue (4): 289-294.doi: 10.11904/j.issn.1002-3070.2016.04.001

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Decorin基因表达对人肺腺癌A549细胞生物学行为的影响

张雪^1,2,王丽¹,燕相伟¹,许庆勇¹

1.哈尔滨医科大学附属肿瘤医院放疗三科(哈尔滨 150086);
2.中航工业哈尔滨市二四二医院肿瘤科

收稿日期:2016-03-16 出版日期:2016-08-10 发布日期:2016-08-19
通讯作者: 许庆勇,E-mail:xuqingyong1976@163.com
作者简介:张雪,女,(1988-),硕士,住院医师,从事肿瘤放疗学的研究
基金资助:
黑龙江省自然科学基金面上项目(D201154)

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

ZHANG Xue^1,2,WANG Li¹,YAN Xiangwei¹,XU Qingyong¹

1.The Third Department of Radiotherapy,The Affiliated Tumor Hospital of Harbin Medical university,Harbin 150086,China;
2.Department of Tumor,The 242th Hospital of Harbin AVIC

Received:2016-03-16 Online:2016-08-10 Published:2016-08-19

摘要/Abstract

摘要： 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LipofectamineTM2000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。

关键词: 核心蛋白聚糖, siRNA干扰, 增殖, 迁移, 侵袭

Abstract: Objective To explore the influences of decorin(DCN)gene down-regulating on biological behavior in human lung adenocarcinoma A549 cell line.Methods Chemically synthesis in vitro targeting DCN-siRNA was transfected into human lung adenocarcinoma A549 cells by LipofectamineTM 2000.The gene expression 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 A549 cells.Real-Time PCR results showed that DCN mRNA expression level significantly decreased,compared with untransfected group,negative control group.Western blot results 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 A549 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-siRNA 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 A549 cells without affecting apoptosis,which provides a new evidence of function for advancing research of lung cancer.

Key words: Decorin gene, siRNA in terference, Proliferation, Migration, Invasion

中图分类号:

R734.2

张雪,王丽,燕相伟,许庆勇. Decorin基因表达对人肺腺癌A549细胞生物学行为的影响[J]. 实用肿瘤学杂志, 2016, 30(4): 289-294.

ZHANG Xue ,WANG Li ,YAN Xiangwei ,XU Qingyong. The influences of Decorin gene on biological behavior in human lung adenocarcinoma A549 cell line[J]. PRACTICAL ONCOLOGY JOURNAL, 2016, 30(4): 289-294.

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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

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