CAJ | 학술논문

目的观察靶向Livin基因反义核酸(ASODN)对肝癌HepG2细胞生长增殖及Livin基因表达的影响.方法应用ASODN、随机寡核苷酸(RODN)分别转染HepG2细胞,采用WST-8法分析细胞增殖抑制效果;反转录-聚合酶链反应(RT-PCR)检测Livin RNA的表达水平;流式细胞术碘化丙锭(PI)单染色检测细胞周期和凋亡.结果靶向Livin反义核酸转染HepG2细胞后,400 nmol/L浓度组作用于肝癌细胞48 h后增殖抑制率达59.5％,与空白对照组和RODN组比较,差异有统计学意义(P＜ 0.05);Livin反义核酸转染HepG2细胞后能明显抑制Livin mRNA的表达,在400 nmol/L浓度组抑制率为52.5％,与空白对照组、RODN组比较,差异有统计学意义(P＜0.05);流式细胞术PI单染空白对照组、RODN组均未出现HepG2细胞亚二倍体峰;100、200、400 nmol/L浓度Livin反义核酸组其亚二倍体百分率分别为13.2％、13.9％、18.9％.结论靶向Livin反义核酸能显著抑制HepG2细胞增殖,下调Livin基因的表达,并诱导HepG2细胞凋亡.
목적 관찰파향Livin기인반의핵산(ASODN)대간암HepG2세포생장증식급Livin기인표체적영향.방법 응용ASODN、수궤과핵감산(RODN)분별전염HepG2세포,채용WST-8법분석세포증식억제효과;반전록-취합매련반응(RT-PCR)검측Livin RNA적표체수평;류식세포술전화병정(PI)단염색검측세포주기화조망.결과 파향Livin반의핵산전염HepG2세포후,400 nmol/L농도조작용우간암세포48 h후증식억제솔체59.5％,여공백대조조화RODN조비교,차이유통계학의의(P＜ 0.05);Livin반의핵산전염HepG2세포후능명현억제Livin mRNA적표체,재400 nmol/L농도조억제솔위52.5％,여공백대조조、RODN조비교,차이유통계학의의(P＜0.05);류식세포술PI단염공백대조조、RODN조균미출현HepG2세포아이배체봉;100、200、400 nmol/L농도Livin반의핵산조기아이배체백분솔분별위13.2％、13.9％、18.9％.결론 파향Livin반의핵산능현저억제HepG2세포증식,하조Livin기인적표체,병유도HepG2세포조망.
Objective To observe the effect of antisense oligonucleotides (ASODN) targeting Livin on proliferation of HepG2 cells and the expression of Livin in vitro.Methods Observe the mechanism of Livin ASODN delivered by LipofectamineTM 2000 on HepG2: (1) The proliferating inhibition of ASODN on HepG2 cells : quantified by WST-8 test in vitro.(2) Test of Livin mRNA expression on HepG2 cells after treated with Livin ASODN by reverse transcriptase-polymerase chain reaction (RT-PCR).(3) Propidine iodide (PI) staining tests to observe these cells cycle changes tested by flow cytometry.Results (1) The inhibition of proliferating can not be seen in the control group and RODN group, while the proliferating inhibition rates of HepG2 cells in ASODN 400 nmoL/L group has reached to 59.5 percent.The difference is statistically significant (P ＜ 0.05).(2) Livin ASODN can significantly inhibit HepG2 cells Livin-mRNA expressions, 400 nmol/L group inhibitory rates have reached to 52.5 percent.Comparing with control group and RODN group, the difference is statistically significant (P ＜ 0.05).(3) PI staining: there was scare hypodiploid in control group and RODN group, while hypodiploid appeared in high dose group of Livin ASODN;the rates of hypodiploid in 100, 200, 400 nmol/L group have reached to 13.2 percent, 13.9 percent and 18.9 percent.Conclusion Livin ASODN can significantly inhibit the proliferating of HepG2 cells, inhibit HepG2 cells Livin-mRNA expressions, and induce these cells to apoptosis.