Zhang Peng, Liu Yi, Wu Zhi-hao, Chen Jun, Chen Gang, Zhou Qing-hua
Thoracic Tumor Center, General Hospital of Tianjin Medical University, Tianjin 300052, China.
Zhonghua Yi Xue Za Zhi. 2009 Dec 1;89(44):3156-9.
To investigate the effects of high mobility group box-1 (HMGB1) silencing upon the invasion and proliferation in human lung cancer cell L9981 by RNA inhibition.
L9981 cells were divided into 3 groups. Group 1 was transfected by HMGB1 small interfering RNA (HMGB1-siRNA group). Group 2 was transfected by negative sequence small interfering RNA (negative control group). Group 3 was blank group. The mRNA and protein of HMGB1 were determined by real-time PCR and Western blotting assay respectively. The proliferation ability was examined by cell viability assay. The growth status of cells was examined by MTT at 24, 48, 72 and 96 h post-transfection. Invasion ability was evaluated by Boyden chamber model.
The relative expression of HMGB1 mRNA of HMGB1-siRNA group (1.0 +/- 0.0) was much lower than the negative control (12.8 +/- 1.3, P < 0.05) and blank groups (12.1 +/- 1.0, P < 0.05). HMGB1 protein expression was also significantly inhibited. Cell viability of HMGB1-siRNA group was much lower than other two groups. MTT indicated the growth of HMGB1-siRNA group was significantly inhibited than other two groups. Boyden chamber model indicated the number of penetrating membrane in HMGB1-siRNA group was less than other two groups.
Down-regulating HMGB1 gene expression by HMGB1 siRNA can inhibit the invasion and proliferation of human lung cancer cell.
通过RNA干扰研究高迁移率族蛋白B1(HMGB1)沉默对人肺癌细胞L9981侵袭和增殖的影响。
将L9981细胞分为3组。第1组用HMGB1小干扰RNA转染(HMGB1-siRNA组)。第2组用阴性序列小干扰RNA转染(阴性对照组)。第3组为空白组。分别采用实时荧光定量PCR和蛋白质印迹法检测HMGB1的mRNA和蛋白。通过细胞活力测定法检测增殖能力。在转染后24、48、72和96小时用MTT检测细胞生长状态。采用Boyden小室模型评估侵袭能力。
HMGB1-siRNA组HMGB1 mRNA的相对表达量(1.0±0.0)明显低于阴性对照组(12.8±1.3,P<0.05)和空白组(12.1±1.0,P<0.05)。HMGB1蛋白表达也受到显著抑制。HMGB1-siRNA组的细胞活力明显低于其他两组。MTT结果显示,HMGB1-siRNA组的细胞生长比其他两组受到显著抑制。Boyden小室模型显示,HMGB1-siRNA组穿过膜的细胞数量少于其他两组。
HMGB1 siRNA下调HMGB1基因表达可抑制人肺癌细胞的侵袭和增殖。