CAJ | 학술논문

目的观察HepG2.2.15细胞在高表达APOBEC3G后,APOBEC3G对细胞生长迁移能力、细胞周期和凋亡现象等生物学行为的影响.方法将真核表达载体pcDNA3.1-APOBEC3G转染入HepG2.2.15细胞中,应用有限稀释法筛选高表达APOBEC3G的HepG2.2.15细胞,并通过Western blot鉴定.应用流式细胞仪检测细胞周期和凋亡;应用噻唑蓝(MTT)比色法检测细胞株生长能力;应用划痕法观察细胞迁徙运动能力.结果 HepG2.2.15-APOBEC3G细胞(pcDNA3.1-APOBEC3G转染)和HepG2.2.15-pcDNA3.1细胞(空载体转染),与HepG2.2.15细胞(空白对照)比较,细胞周期无明显变化[各期细胞所占比例分别为:G0/G1:(64.27±1.26)%比(64.18±1.24)%比(64.09±1.30)%,P＞0.05;S:(23.16±1.38)%比(22.67±1.41)%比(23.27±1.43)%,P＞0.05;G2/M:(11.36±1.26)%比(11.84±1.31)%比(11.37±1.22)%,P＞0.05],未见细胞凋亡现象,细胞的生长增殖无明显变化(P＞0.05),细胞迁徙运动能力无明显变化[24h及48 h细胞迁移抑制率分别为:(7.5±3.7)%比(10.2±5.5)%,P＞0.05;(13.4±6.5)%比(17.8±9.2)%,P＞0.05].结论高表达APOBEC3G对HepG2.2.15细胞生物学行为无明显影响.
목적 관찰HepG2.2.15세포재고표체APOBEC3G후,APOBEC3G대세포생장천이능력、세포주기화조망현상등생물학행위적영향.방법 장진핵표체재체pcDNA3.1-APOBEC3G전염입HepG2.2.15세포중,응용유한희석법사선고표체APOBEC3G적HepG2.2.15세포,병통과Western blot감정.응용류식세포의검측세포주기화조망;응용새서람(MTT)비색법검측세포주생장능력;응용화흔법관찰세포천사운동능력.결과 HepG2.2.15-APOBEC3G세포(pcDNA3.1-APOBEC3G전염)화HepG2.2.15-pcDNA3.1세포(공재체전염),여HepG2.2.15세포(공백대조)비교,세포주기무명현변화[각기세포소점비례분별위:G0/G1:(64.27±1.26)%비(64.18±1.24)%비(64.09±1.30)%,P＞0.05;S:(23.16±1.38)%비(22.67±1.41)%비(23.27±1.43)%,P＞0.05;G2/M:(11.36±1.26)%비(11.84±1.31)%비(11.37±1.22)%,P＞0.05],미견세포조망현상,세포적생장증식무명현변화(P＞0.05),세포천사운동능력무명현변화[24h급48 h세포천이억제솔분별위:(7.5±3.7)%비(10.2±5.5)%,P＞0.05;(13.4±6.5)%비(17.8±9.2)%,P＞0.05].결론 고표체APOBEC3G대HepG2.2.15세포생물학행위무명현영향.
Objective To observe the effects of APOBEC3G on biological behaviors, including cell growth and migration ability, cell cycle and apoptosis of HepG2. 2. 15 cells after highly expressing APOBEC3G. Methods Eukaryotic expression vector pcDNA3. 1-APOBEC3G was transfected into HepG2.2.15 cells. HepG2.2.15 cells highly expressing APOBEC3G protein were screened by using limiting dilution method, and identified by using Western blotting. The changes of cell cycle and apoptosis were detected by using flow cytometry (FCM). The proliferation of cells was analyzed by MTT assay. The migration and movement of cells was determined by rowing trace. Results There were no significant changes in cell cycle among HepG2.2. 15-APOBEC3G cells (transfected with pcDNA3. 1-APOBEC3G),HepG2.2.15-pcDNA3.1 cells (transfected with blank vector) and HepG2.2.15 cells (control)[the proportions of each phase of cells were as follows: for G0/G1: (64.27 ± 1.26)% vs (64.18 ± 1.24)% vs (64.09 ±1.30)%,P＞0.05; for S: (23.16 ±1.38)% vs (22.67 ±1.41)% vs (23.27 ±1.43)%,P＞0.05; for G2/M: (11.36±1.26)% vs (11.84±1.31)% vs (11.37 ±1.22)%,P＞0.05, respectively], apoptosis (P ＞ 0.05), cell growth (P ＞ 0.05 ), and the ability of cell migration[the inhibition rate of cell migration at 24 and 48 h was (7.5 ±3.7)% vs (10.2 ±5.5)% ,P＞0.05; (13.4 ±6.5)%vs ( 17.8 ± 9.2 ) %, P ＞ 0.05, respectively]. Conclusion Highly-expressed APOBEC3G has no obvious effects on biological behaviors of HepG2.2.15 cells.