CAJ | 학술논문

目的观察以基质金属蛋白酶为靶点的肿瘤靶向性细胞穿膜肽介导小干扰RNA(siRNA)进入肝癌并对肝癌细胞SMMC-7721起一定抑制作用.方法以体外培养的肝癌细胞SMMC-7721为研究对象,将细胞分为实验组和对照组,实验组加入siRNA质粒/穿膜肽复合物,对照组加入空质粒/穿膜肽复合物,培养48 h后收集两组细胞,流式细胞仪检测各组细胞的细胞周期,逆转录-聚合酶链反应(RT-PCR)检测各组细胞人端粒酶逆转录酶(hTERT)基因的表达水平,噻唑蓝(MTT)法检测SiRNA质粒/穿膜肽复合物对细胞增殖的抑制作用.结果实验组与对照组比较G1期细胞所占比例增多(实验组75.14％、对照组62.55％),G2期细胞比例减少(实验组11.39％、对照组12.14％)、S期细胞比例减少(实验组13.47％、对照组25.31％),说明经穿膜肽质粒复合物处理后细胞被阻滞在G1期,实验组细胞hTERT mRNA的表达量约为未对照组的74％,即实验组mRNA表达水平下调约26％.肝癌细胞经穿膜肽复合物处理48 h后MTT法测细胞增殖抑制率为34.82％.结论肿瘤靶向性细胞穿膜肽可介导siRNA进入肝癌细胞并可对肝癌细胞起一定的抑制作用.
목적 관찰이기질금속단백매위파점적종류파향성세포천막태개도소간우RNA(siRNA)진입간암병대간암세포SMMC-7721기일정억제작용.방법 이체외배양적간암세포SMMC-7721위연구대상,장세포분위실험조화대조조,실험조가입siRNA질립/천막태복합물,대조조가입공질립/천막태복합물,배양48 h후수집량조세포,류식세포의검측각조세포적세포주기,역전록-취합매련반응(RT-PCR)검측각조세포인단립매역전록매(hTERT)기인적표체수평,새서람(MTT)법검측SiRNA질립/천막태복합물대세포증식적억제작용.결과 실험조여대조조비교G1기세포소점비례증다(실험조75.14％、대조조62.55％),G2기세포비례감소(실험조11.39％、대조조12.14％)、S기세포비례감소(실험조13.47％、대조조25.31％),설명경천막태질립복합물처리후세포피조체재G1기,실험조세포hTERT mRNA적표체량약위미대조조적74％,즉실험조mRNA표체수평하조약26％.간암세포경천막태복합물처리48 h후MTT법측세포증식억제솔위34.82％.결론 종류파향성세포천막태가개도siRNA진입간암세포병가대간암세포기일정적억제작용.
Objective To observe the inhibitory effect of small interfering RNA (siRNA) mediated by tumor target cell penetrating peptides (CPPS) targeting metalloproteinase on hepatoma cell line SMMC-7721.Methods SMMC-7721 cells were cultured,and divided into experimental group and control group.In experimental group,siRNA/CCPS complex was added,and in control group,silencer negative control/CCPS was added.The cells were harvested after culture for 48 h.Flow cytometry was use to examine the cell cycle.The human telomerase reverse transcriptase (hTERT) gene expression level was detected by using quantitative polymerase chain reaction (PCR).Methyl thiazol tetrazolium (MTT) assay was used to meassure the inhibitory effect of siRNA/CCPS complex on SMMC-7721 cells.Results As compared with control group,the proportion of cells in G1 phase in experimental group was increased (75.14％ vs.62.55％),while that in G2 phase in experimental group decreased (11.39％ vs.12.14％),and that in the S phase also reduced (13.47％ vs.25.31％).Treatment of CPPS plasmid complexes could arrest the cells in the G1 phase,and the expression quantity of hTERT mRNA in experimental group was 74％ of that in control group.After treatment with CPPS plasmid complexes for 48 h,the proliferation inhibition rate of SMMC-7721 cells was 34.82％.Conclusion The CCPS can be a transmitter to transfect siRNA into hepatoma cells and suppress hepatoma cells to some extent.