长链非编码RNA PTENP1基因抑制肝癌细胞增殖和迁移  

Long noncoding RNA PTENP1 inhibits proliferation and migration of hepatocellular carcinoma cells

作　　者：熊志勇[1] 姚志成[2] 范伟明[1] 李明亮[3] 胡昆鹏[2] 徐见亮[1] 钟跃思[1] 许瑞云[1] 邓美海[1] 

机构地区：[1]中山大学附属第三医院肝胆外科,广州510630 [2]中山大学附属第三医院普通外科,广州510630 [3]中山大学附属第三医院重症监护室,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2016年第2期119-123,共5页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金(8157111144);广东省自然科学基金(S2013010016015);广东省科技计划项目(2013B01040413;2014A020212122);广州市科技计划项目(1563000226)

摘　　要：目的探讨长链非编码RNA(lnc RNA)PTEN假基因1(PTENP1)对肝癌细胞增殖和迁移能力的影响及机制。方法合成表达PTENP1的慢病毒载体,分别将LV003-GFP-PTENP1病毒和LV003-GFP空载病毒感染肝癌细胞BEL-7404,构建稳定表达PTENP1肝癌细胞的实验组和对照组。采用CCK-8法和平板克隆实验检测两组肝癌细胞增殖能力和克隆形成能力,划痕实验检测肝癌细胞迁移能力,Western blot法检测p44/42丝裂原活化蛋白激酶(MAPK)、p38 MAPK蛋白表达情况。结果实验组细胞48、72 h的吸光度值A450分别为1.4±0.3、2.3±1.1,明显低于对照组的3.2±1.7、3.4±1.1(t=-5.78,-4.23;P<0.05)。实验组细胞克隆形成数目为(55±12)个,明显低于对照组的(154±45)个(t=-3.98,P<0.05)。实验组划痕实验的细胞迁移率为(21.7±2.6)%,明显低于对照组的(57.7±4.9)%(t=-8.34,P<0.05)。Western blot检测显示,实验组p44/42 MAPK、p38MAPK蛋白表达较对照组明显下降。结论 lnc RNA PTENP1可抑制肝癌细胞的增殖和迁移,其机制可能是通过MAPK信号通路调控。Objective To investigate the effect and mechanism of long noncoding RNA (lncRNA) PTEN pseudogene 1 (PTENP1) on the proliferation and migration of hepatocellular carcinoma (HCC) cells. Methods Lentiviral vectors expressing PTENP1 were constructed. HCC cells BEL-7404 were infected with LV003-GFP-PTENP1 and control vectors LV003-GFP. BEL-7404 cells stably expressing PTENP1 were constructed and the experimental and control groups were established. The proliferation and clone formation abilities of HCC cells in two groups were detected by CCK-8 assay and clonogenic assay. The migration ability of HCC cells was detected by wound healing assay. The expression of p44/42 mitogen-activated protein kinase (MAPK) and p38 MAPK proteins were detected by Western blot. Results The absorbance values A450 of the cells at 48 and 72 h in the experimental group were 1.4±0.3 and 2.3±1.1, signiifcantly lower compared with 3.2±1.7 and 3.4±1.1 in the control group (t=-5.78,-4.23;P<0.05). The number of cell clone&nbsp;formation in the experimental group was 55±12, signiifcantly less than 154±45 in the control group (t=-3.98, P<0.05). The percentage of cell migration in the experimental group was (21.7±2.6)%, signiifcantly lower than (57.7±4.9)%in the control group (t=-8.34, P<0.05). Western blot revealed that the expression of p44/42 MAPK and p38 MAPK proteins in the experimental group was significantly down-regulated compared with those in the control group. Conclusion lncRNA PTENP1 can inhibit the proliferation and migration of HCC cells probably through regulating MAPK signaling pathway.

关 键 词：核糖核酸酶类 长链非编码RNA PTENP1 肝肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]