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1(LRIG1)基因过表达对人肝癌细胞株Hep-G2体内外恶性表型的影响并探讨其作用机制.方法使用10％胎牛血清(FBS)的DMEM(改良伊戈尔培养基)培养基对Hep2进行培养48 h后,分别进行不转染,转染pEGFP-N1和转染pEGFP-N1-LRIG1的处理,相对应分为未处理组、pEGFP-N1空载体组和pEGFP-N1-LRIG1实验组.再采用细胞计数试剂盒(CCK-8)比色法、Transwell法、实时荧光定量聚合酶链反应(Real-time PCR)及Western blot等方法分析LRIG1基因过表达对肝癌细胞株Hep-G2生长增殖和侵袭能力的影响,以及胞膜上表皮生长因子受体(EGFR)的mRNA与蛋白水平的变化.结果 CCK-8结果显示,未处理组、pEGFP-N1空载体组、pEGFP-N1-LRIG1质粒组在24、48、72h作用时间处的吸光度值分别为:0.968±0.050、0.924±0.070、0.552±0.060;0.975±0.120、0.864±0.200、0.509±0.130;0.983±0.180、0.803±0.140、0.465±0.230；与未处理组和空载体组比较,LRIG1对Hep-G2细胞有明显抑制效应(P＜0.05)；细胞侵袭实验结果表明,LRIG1过表达组、未处理及空载体组侵袭实验透膜细胞数分别为(6.24±1.52)个、(15.19±1.32)个和(14.85±1.24)个(P＜0.05)；与未处理组和空载体组比较,LRIG1能显著抑制Hep-G2细胞的侵袭能力(P＜ 0.05)；Real-time PCR及Western blot实验结果显示,LRIG1过表达组能显著抑制EGFR在Hep-G2细胞中mRNA和蛋白水平的表达(P＜0.05).结论 LRIG1可能通过抑制EGFR的表达来抑制Hep-G2细胞在体内外的增殖.
목적 관찰부함량안산중복서렬화면역구단백양결구역기인1(LRIG1)기인과표체대인간암세포주Hep-G2체내외악성표형적영향병탐토기작용궤제.방법 사용10％태우혈청(FBS)적DMEM(개량이과이배양기)배양기대Hep2진행배양48 h후,분별진행불전염,전염pEGFP-N1화전염pEGFP-N1-LRIG1적처리,상대응분위미처리조、pEGFP-N1공재체조화pEGFP-N1-LRIG1실험조.재채용세포계수시제합(CCK-8)비색법、Transwell법、실시형광정량취합매련반응(Real-time PCR)급Western blot등방법분석LRIG1기인과표체대간암세포주Hep-G2생장증식화침습능력적영향,이급포막상표피생장인자수체(EGFR)적mRNA여단백수평적변화.결과 CCK-8결과현시,미처리조、pEGFP-N1공재체조、pEGFP-N1-LRIG1질립조재24、48、72h작용시간처적흡광도치분별위:0.968±0.050、0.924±0.070、0.552±0.060;0.975±0.120、0.864±0.200、0.509±0.130;0.983±0.180、0.803±0.140、0.465±0.230；여미처리조화공재체조비교,LRIG1대Hep-G2세포유명현억제효응(P＜0.05)；세포침습실험결과표명,LRIG1과표체조、미처리급공재체조침습실험투막세포수분별위(6.24±1.52)개、(15.19±1.32)개화(14.85±1.24)개(P＜0.05)；여미처리조화공재체조비교,LRIG1능현저억제Hep-G2세포적침습능력(P＜ 0.05)；Real-time PCR급Western blot실험결과현시,LRIG1과표체조능현저억제EGFR재Hep-G2세포중mRNA화단백수평적표체(P＜0.05).결론 LRIG1가능통과억제EGFR적표체래억제Hep-G2세포재체내외적증식.
Objective To investigate the effect of high mobility group leucine-rich repeats and immunoglobulin-like domain1 gene(LRIG1) on the proliferative activity of human hepatoma cell line Hep-G2 in vivo and in vitro and its potential action mechanism.Methods The Hep-G2 cells,cultured for 24 h by DMEM supplemented with 10％ fetal bovine serum (FBS),were treated with none,pEGFP-N1 and pEGFP-N1-LRIG1.Cell proliferation was observed by cell counting kit-8 (CCK-8) analysis.Transwell invasion assay was used to monitor the changes of Hep-G2 cell malignant phenotypes.Changes of LRIG1 and epidermal growth factor receptor (EGFR) expression at mRNA and protein levels were measured by realtime polymerase chain reaction (PCR) and Western blotting,respectively.Results The absorbance (A) values at 24,48 and 72 h in control group,pEGFP-N1-LRIG1 group and pEGFP-N1 group were 0.968 ±0.050,0.924±0.070,0.552 ±0.060; 0.975±0.120,0.864 ±0.200,0.509±0.130; 0.983 ±0.180,0.803 ±0.140,0.465 ±0.230.As compared with control group and pEGFP-N1 group,proliferation of Hep-G2 cells was significantly inhibited in pEGFP-N1-LRIG1 group (P ＜ 0.05).The number of invasive cells in pEGFP-N1-LRIG1 group,control group and pEGFP-N1 group was (6.24 ± 1.52),(15.19 ± 1.32) and (14.85 ± 1.24),respectively.As compared with control group and pEGFP-N1 group,the invasive ability of Hep-G2 cells was significantly inhibited in pEGFP-N1 group (P ＜ 0.05).LRIG1 could significantly suppress the expression of epidermal growth factor receptor (EGFR) mRNA and protein in Hep-G2 cells in vitro (P ＜ 0.05).Conclusion LRIG1 could promote malignant phenotypes of Hep-G2 cells probably by inhibiting the expression of EGFR mRNA and protein.