CAJ | 학술논문

目的构建人肠凝集素-1( ITLN-1)基因的真核表达载体,观察其在胃癌细胞中的表达.方法采用逆转录聚合酶链反应(RT-PCR)法扩增ITLN-1 cDNA,定向克隆至真核表达载体pEGFP-N1,测序鉴定；在脂质体的介导下,重组质粒瞬时转染人胃癌细胞株SGC-7901 72 h后,Western blot法检测细胞培养上清中ITLN-1表达,四甲基偶氮唑盐比色(MTT)法、Transwell小室实验检测癌细胞增殖、侵袭活性.结果真核表达载体pEGFP-ITLN-1转染SGC-7901细胞72 h后,ITLN-1 蛋白表达上调5.72倍(P＜0.01),细胞增殖活性降低52.8％ (P＜ 0.01),细胞侵袭能力下调58.1％ (P＜0.01).结论成功构建人ITLN-1真核表达载体,转染胃癌细胞过表达后,能抑制癌细胞的增殖和侵袭活性.
목적 구건인장응집소-1( ITLN-1)기인적진핵표체재체,관찰기재위암세포중적표체.방법 채용역전록취합매련반응(RT-PCR)법확증ITLN-1 cDNA,정향극륭지진핵표체재체pEGFP-N1,측서감정；재지질체적개도하,중조질립순시전염인위암세포주SGC-7901 72 h후,Western blot법검측세포배양상청중ITLN-1표체,사갑기우담서염비색(MTT)법、Transwell소실실험검측암세포증식、침습활성.결과 진핵표체재체pEGFP-ITLN-1전염SGC-7901세포72 h후,ITLN-1 단백표체상조5.72배(P＜0.01),세포증식활성강저52.8％ (P＜ 0.01),세포침습능력하조58.1％ (P＜0.01).결론 성공구건인ITLN-1진핵표체재체,전염위암세포과표체후,능억제암세포적증식화침습활성.
Objective To construct the eukaryotic expression vector of human intelectin-1 (ITLN-1)and observe its expression in gastric cancer cells.Methods ITLN-1 cDNA was amplified by reverse transcription polymerase chain reaction ( RT-PCR),inserted into eukaryotic vector pEGFP-N1,and validated by nucleic acid sequencing.Under the induction of Lipofectamine 2000,the recombinant was transfected into SGC-7901 cells for 72 hours.The expression level of ITLN-1 in medium supematant was detected by western blotting.2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetry and transwell assay were applied to measure the in vitro proliferation and invasion of cancer cells.Results Seventy-two hours post-transfection of pEGFP-ITLN-1 into SGC-7901 cells,the ITLN-1 protein level was upregulated by 5.72 times (P ＜ 0.01 ).Meanwhile,the cellular proliferation and invasive activity were downregulated by52.8％ (P＜0.01) and 58.1％ (P＜0.01),respectively.Conclusion The expression vector of ITLN-1 was successfully constructed,and induced the ITLN-1 overexpression in gastric cancer cells,resulting in decrease of proliferation and invasion of cancer cells.