CAJ | 학술논문

目的：探讨 E2F -1过表达对胃癌 SGC -7901细胞增殖、生长和细胞周期的影响及其分子机制。方法采用携带 E2F -1基因的重组慢病毒颗粒（LV - E2F -1- GFP）感染人胃癌 SGC -7901细胞（LV - E2F -1-GFP 组）；以对照慢病毒颗粒（LV - GFP）感染人胃癌 SGC -7901细胞作为阴性对照（LV - GFP 组）；空白对照组常规培养胃癌 SGC -7901细胞，不作任何处理。分别采用 CCK -8法检测细胞的增殖活力，流式细胞仪检测各组细胞周期的分布，RT - PCR 和 Western blot 技术检测细胞中 Skp2、Bax、Bcl -2、CylinD1和 Survivin 基因 mRNA 和蛋白的表达。结果与 LV - GFP 组和空白对照组比较，LV - E2F -1- GFP 组人胃癌 SGC -7901细胞增殖活力明显降低（P <0.05），G0/ G1期所占比例上升（P <0.05），Skp2、Bcl -2、CylinD1和 Survivin 基因的 mRNA 和蛋白的表达降低（P <0.05），Bax 基因的 mRNA 和蛋白的表达上调（P <0.05），而 LV - GFP 组和空白对照组比较，差异无统计学意义（P >0.05）。结论慢病毒介导 E2F -1基因过表达抑制胃癌细胞增殖和生长，使细胞周期停滞在 G0/ G1期，其机制可能与 E2F -1基因过表达使胃癌细胞 Skp2、Survivin、Bcl -2、CylinD1基因表达下调和 Bax 基因表达上调有关。
목적：탐토 E2F -1과표체대위암 SGC -7901세포증식、생장화세포주기적영향급기분자궤제。방법채용휴대 E2F -1기인적중조만병독과립（LV - E2F -1- GFP）감염인위암 SGC -7901세포（LV - E2F -1-GFP 조）；이대조만병독과립（LV - GFP）감염인위암 SGC -7901세포작위음성대조（LV - GFP 조）；공백대조조상규배양위암 SGC -7901세포，불작임하처리。분별채용 CCK -8법검측세포적증식활력，류식세포의검측각조세포주기적분포，RT - PCR 화 Western blot 기술검측세포중 Skp2、Bax、Bcl -2、CylinD1화 Survivin 기인 mRNA 화단백적표체。결과여 LV - GFP 조화공백대조조비교，LV - E2F -1- GFP 조인위암 SGC -7901세포증식활력명현강저（P <0.05），G0/ G1기소점비례상승（P <0.05），Skp2、Bcl -2、CylinD1화 Survivin 기인적 mRNA 화단백적표체강저（P <0.05），Bax 기인적 mRNA 화단백적표체상조（P <0.05），이 LV - GFP 조화공백대조조비교，차이무통계학의의（P >0.05）。결론만병독개도 E2F -1기인과표체억제위암세포증식화생장，사세포주기정체재 G0/ G1기，기궤제가능여 E2F -1기인과표체사위암세포 Skp2、Survivin、Bcl -2、CylinD1기인표체하조화 Bax 기인표체상조유관。
Objective To investigate the effect of E2F -1 overexpression on the proliferation,growth and cell cy-cle of human gastric cancer cell line SGC -7901 and the molecular mechanism. Methods Humans gastric cancer SGC -7901 cells were infected by E2F - 1 recombinant lentiviral particles(LV - E2F - 1 - GFP),as experimental group(LV -E2F - 1 - GFP group),while control lentiviral particles(LV - GFP)infected human gastric cancer SGC - 7901 cells,as a negative control group(LV - GFP group)and blank control group was cultured conventionally without any treatment. CCK - 8 were used to detect cell proliferation activity,while flow cytometry was used for assessment of cell cycle distribu-tion. Semi - quantitative reserve transcription - polymerase chain reaction(RT - PCR)and Western blot were applied for measurement of mRNA and protein expression of Skp2,Bax,Bcl - 2,CylinD1 and Survivin. Results Compared with LV - GFP group and blank control group,significant lower proliferation activity(P < 0. 05),increased G0 / G1 phase pro-portion(P < 0. 05),down - regulation of mRNA and protein of Skp2,Bcl - 2,CylinD1 and Survivin(P < 0. 05),up -regulation of Bax mRNA and protein(P < 0. 05),were revealed LV - E2F - 1 - GFP group. Furthermore,there was no significant difference between LV - GFP group and blank control group(P > 0. 05). Conclusion Lentivirus - mediated overexpression of E2F - 1 induces growth inhibition of cancer cells,and cell cycle arrest at G0 / G1 phase;probably via up - regulation of Bax and down - regulation of Skp2,Survivin,Bcl - 2 and CylinD1.