CAJ | 학술논문

目的观察富含亮氨酸重复序列和免疫球蛋白样结构域基因3( LRIG3)在3种肝癌细胞株HepG-2、LM3和7721中的表达,探讨LRIG3基因对肝癌细胞株HepG-2、LM3和7721的细胞周期、侵袭能力和凋亡的影响.方法将过表达的LRIG3质粒和空白质粒经脂质体法分别转染肝癌细胞株HepG-2、LM3和7721中,相对应分为实验组和对照组,采用逆转录-聚合酶链反应(RT-PCR)和Western blot法检测LRIG3表达水平变化,采用流式细胞仪、Transwell法及原位末端转移酶标记(TUNEL)等方法对细胞株的生物学行为进行分析.结果与对照组比较,实验组HepG-2、LM3和7721细胞中LRIG3 mRNA水平分别上升78.9％、67.8％和59.6％,蛋白表达水平分别升高91.2％、79.9％和65.7％,差异有统计学意义(P＜0.05)；实验组HepG-2和LM3细胞中穿出细胞数量均明显少于对照组,差异有统计学意义(P＜0.05)；TUNEL染色结果显示对照组HepG-2细胞凋亡率为23.4％,实验组为35.7％.对照组LM3细胞凋亡率为21.6％,实验组为39.5％.对照组7721细胞凋亡率为20.2％,实验组为30.5％,对照组与实验组间凋亡率差异有统计学意义(P＜0.05).实验组HepG-2和LM3细胞中S期与G2/M期细胞数之和低于对照组,差异有统计学意义(P＜0.05),但7721细胞与对照组比较差异无统计学意义(P＞0.05).结论 LRIG3基因过表达能阻断细胞周期于S期和G2/M期,降低肝癌细胞的侵袭能力,加速细胞凋亡.
목적 관찰부함량안산중복서렬화면역구단백양결구역기인3( LRIG3)재3충간암세포주HepG-2、LM3화7721중적표체,탐토LRIG3기인대간암세포주HepG-2、LM3화7721적세포주기、침습능력화조망적영향.방법 장과표체적LRIG3질립화공백질립경지질체법분별전염간암세포주HepG-2、LM3화7721중,상대응분위실험조화대조조,채용역전록-취합매련반응(RT-PCR)화Western blot법검측LRIG3표체수평변화,채용류식세포의、Transwell법급원위말단전이매표기(TUNEL)등방법대세포주적생물학행위진행분석.결과 여대조조비교,실험조HepG-2、LM3화7721세포중LRIG3 mRNA수평분별상승78.9％、67.8％화59.6％,단백표체수평분별승고91.2％、79.9％화65.7％,차이유통계학의의(P＜0.05)；실험조HepG-2화LM3세포중천출세포수량균명현소우대조조,차이유통계학의의(P＜0.05)；TUNEL염색결과현시대조조HepG-2세포조망솔위23.4％,실험조위35.7％.대조조LM3세포조망솔위21.6％,실험조위39.5％.대조조7721세포조망솔위20.2％,실험조위30.5％,대조조여실험조간조망솔차이유통계학의의(P＜0.05).실험조HepG-2화LM3세포중S기여G2/M기세포수지화저우대조조,차이유통계학의의(P＜0.05),단7721세포여대조조비교차이무통계학의의(P＞0.05).결론 LRIG3기인과표체능조단세포주기우S기화G2/M기,강저간암세포적침습능력,가속세포조망.
Objective To explore the effect of overexpression of leucine-rich repeats and immunoglobin-like domains protein 3 ( LRIG3 ) gene on the cycle,invasion and apoptosis of hepatocellular carcinoma cell lines HepG-2,LM3 and 7721.Methods The plasmids of over-expressed LRIG3 and the blank plasmid serving as control were transduced into HepG-2,LM3 and 7721 cells.The expression levels of LRIG3 mRNA and protein were detected by using reverse transcription-polymerase chain reaction (RTPCR) and Western blotting,and the changes in the cell cycle were determined by flow cytometry.The invasive capability was measured by Transwell assay,and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method was used to measure apoptosis.Results As compared with the control group,the mRNA levels of LRIG3 were increased by 78.9％,67.8％ and 59.6％ respectively,and their protein levels were increased by 91.2％,79.9％ and 65.7％ in HepG-2,LM3 and 7721 cells of experiment group,respectively ( P ＜0.05).The average number of HepG-2,LM3 and 7721 cells over-expressing LRIG3 passing through the inserted filter was decreased as compared with control group ( P ＜ 0.05 ).The number of cells in S + G2/M phase in HepG-2 and LM3 cells of experiment group was significantly reduced as compared with control group (P ＜0.05).The apoptosis rate of HepG-2,LM3 and 7721 cells was 35.7％,39.5％ and 30.5％ respectively in experiment groups,and that was 23.4％,21.6％ and 20.2％respectively in control groups.There was significant difference between experiment group and control group ( P ＜ 0.05 ).Conclusion The over-expression of LRIG3 gene could arrest the cell in S and G2/M phases,reduce the invasive capability of HCC,and accelerate apoptosis of HCC.