中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2015年
6期
410-414
,共5页
黄山%刘柯%朱继业%杨琦%韦小波%王琪%吴飞翔%黎乐群
黃山%劉柯%硃繼業%楊琦%韋小波%王琪%吳飛翔%黎樂群
황산%류가%주계업%양기%위소파%왕기%오비상%려악군
趋化因子,CXCL-1%肝癌细胞%稳定转染%增殖%侵袭
趨化因子,CXCL-1%肝癌細胞%穩定轉染%增殖%侵襲
추화인자,CXCL-1%간암세포%은정전염%증식%침습
Chemokine,CXCL-1%Hepatocellular carcinoma cells%Stable transfection%Proliferation%Invasion
目的 建立慢病毒载体介导稳定转染趋化因子CXCL-1的MHCC97-L肝癌细胞株,鉴定转染细胞中目的基因的表达并观察转染CXCL-1对MHCC97-L细胞增殖、迁移、侵袭能力及细胞周期的影响.方法 慢病毒载体介导CXCL-1基因稳定转染MHCC97-L肝癌细胞,qRT-PCR、蛋白印迹方法分别检测转染后细胞中CXCL-1在mRNA和蛋白水平的表达;MTT试验、克隆形成试验,Transwell小板细胞穿膜试验分别检测转染后细胞增殖能力、迁移能力及侵袭能力的变化.结果 成功建立了稳定转染CXCL-1的MHCC97-L肝癌细胞株.转染后细胞高表达CXCL-1基因.转染后MHCC97-L细胞增殖能力显著增强(P<0.05),侵袭能力显著增强(P<0.05),迁移能力变化则无统计学差异(P>0.05).转染后细胞株中G1期细胞比例显著减少(P<0.05).结论 成功构建稳定转染CXCL-1的肝癌细胞株,CXCL-1可增强肝癌细胞的体外增殖和侵袭能力.
目的 建立慢病毒載體介導穩定轉染趨化因子CXCL-1的MHCC97-L肝癌細胞株,鑒定轉染細胞中目的基因的錶達併觀察轉染CXCL-1對MHCC97-L細胞增殖、遷移、侵襲能力及細胞週期的影響.方法 慢病毒載體介導CXCL-1基因穩定轉染MHCC97-L肝癌細胞,qRT-PCR、蛋白印跡方法分彆檢測轉染後細胞中CXCL-1在mRNA和蛋白水平的錶達;MTT試驗、剋隆形成試驗,Transwell小闆細胞穿膜試驗分彆檢測轉染後細胞增殖能力、遷移能力及侵襲能力的變化.結果 成功建立瞭穩定轉染CXCL-1的MHCC97-L肝癌細胞株.轉染後細胞高錶達CXCL-1基因.轉染後MHCC97-L細胞增殖能力顯著增彊(P<0.05),侵襲能力顯著增彊(P<0.05),遷移能力變化則無統計學差異(P>0.05).轉染後細胞株中G1期細胞比例顯著減少(P<0.05).結論 成功構建穩定轉染CXCL-1的肝癌細胞株,CXCL-1可增彊肝癌細胞的體外增殖和侵襲能力.
목적 건립만병독재체개도은정전염추화인자CXCL-1적MHCC97-L간암세포주,감정전염세포중목적기인적표체병관찰전염CXCL-1대MHCC97-L세포증식、천이、침습능력급세포주기적영향.방법 만병독재체개도CXCL-1기인은정전염MHCC97-L간암세포,qRT-PCR、단백인적방법분별검측전염후세포중CXCL-1재mRNA화단백수평적표체;MTT시험、극륭형성시험,Transwell소판세포천막시험분별검측전염후세포증식능력、천이능력급침습능력적변화.결과 성공건립료은정전염CXCL-1적MHCC97-L간암세포주.전염후세포고표체CXCL-1기인.전염후MHCC97-L세포증식능력현저증강(P<0.05),침습능력현저증강(P<0.05),천이능력변화칙무통계학차이(P>0.05).전염후세포주중G1기세포비례현저감소(P<0.05).결론 성공구건은정전염CXCL-1적간암세포주,CXCL-1가증강간암세포적체외증식화침습능력.
Objective To establish a MHCC97-L cell lines which stably express CXCL-1 by using lentiviral transfection system and investigate the effect of CXCL1 on the cell proliferation,migration and invasion as well as the cell cycle.Methods The expression levels of mRNA and protein in MHCC97-L cell line with the CXCL1 lentivirus-mediated transfection were evaluated by qRT-PCR and western blot,respectively.We conducted MTT,clone formation and Transwell assay to examine the changes of cell proliferation,migration and invasion in the transfected cells.Results A stably transfected cell line was successfully established.Cell proliferation (P < 0.05) and invasion (P < 0.05) of MHCC97-L cells transfected with CXCL-1 gene tended to be significantly enhanced whilst nothing seemed to alter in cell migration (P > 0.05).The percentage of transfected cells in the G1 stage declined significantly (P < 0.05).Conclusion A stable cell line transfected with CXCL1 was established,and CXCL1 overexpression could promote the proliferation and invasion of the hepatocellular carcinoma cells in vitro.