survivin基因表达对肺鳞状细胞癌细胞侵袭的影响

Influence of survivin gene expression on invasion of human squamous carcinoma cell

目的探讨survivin基因对肺鳞状细胞癌细胞侵袭能力的影响及作用机制。方法应用survivin基因小干扰RNA(siRNA)转染处理人肺鳞状细胞癌细胞后,用荧光实时定量PCR和Western blot检测survivin mRNA和survivin蛋白表达,用软琼脂集落培养试验和Boyden小室模型试验检测癌细胞的锚着不依赖性增殖和侵袭能力,用琼脂糖凝胶电泳和细胞凋亡原位检测试剂盒检测癌细胞失巢凋亡情况。结果转染组癌细胞survivin mRNA和survivin蛋白表达明显受抑制,且与时间和浓度相关。与对照组比较,转染组细胞形成的软琼脂集落数和穿膜细胞数明显减少,细胞出现明显凋亡现象,并出现明显DNA条带。结论survivin基因siRNA转染可明显抑制肺鳞状细胞癌细胞的锚着不依赖性增殖和恶性侵袭,其机制可能与诱导失巢凋亡有关。

Objective To study the effects of survivin gene on invasion and it＇s mechanism of human lung squamous carcinoma cell. Methods After human lung squamous cell carcinoma NCI-H520 cell were transfected by survivin small interfering RNA（ siRNA）, we studied the expression of survivin mRNA and protein using real-time PCR and Western bloting, respectively. Anchorage-independent growth was measured by elon formation in soft agar, and invasusing hoyden chamber model, and anoikis of cancer cell was measured by agarose gel electrophoresis and DNA fragmentation assay and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay. Results The levels of mRNA and protein of survivin in group transfectd with siRNA were decceased obviously in a time-dependent and dose-dependent manner. Compared with control group, both colony formation in soft agar and cells traversed membrane were reduced significantly, and cell apoptosis and DNA strap were appeared. Conclusion Survivin gene transfected by siRNA can resist the anchorage-independent growth and malignant invasion, which may be related with anoikis of cancer cell.