CAJ | 학술논문

目的:探讨应用RNAi技术下调bcl-2基因表达后对人黑素瘤Mi4细胞株生长的影响.方法:采用人黑素瘤M14细胞株,利用人工合成的bel-2靶向小分子干扰RNA(small interferenceRNA,siRNA)转染细胞,MTT法测细胞增殖情况.结果:免疫组化可见25例实验组细胞爬片均表达bcl-2,表达强度弱阳性,25例空白对照组和阴性对照均表达bel-2,表达强度强阳性,实验组细胞与空白对照组和阴性对照组相比bcl-2表达明显减低,差异有统计学意义(P<0.05),空白对照组和阴性对照组bcl-2表达差异无统计学意义(P>0.05),MTT法测定显示:转染48h后,实验组7组细胞平均吸光度OD值0.384±0.026,空白对照组7组细胞平均吸光度OD值0.590±0.032,阴性对照组7组细胞平均吸光度OD 值0.541±0.034,实验组细胞吸光度降低,与空白对照组及阴性对照组相比差异有统计学意义(P<0.05),而空白对照组和阴性对照组两组间差异无统计学意义(P>0.05),实验组细胞增殖抑制率34.91%,阴性对照组细胞增殖抑制率8.30%,差异有统计学意义(P<0.05),显示实验细胞增殖减慢.结论:Bel-2RNAi能显著降低人黑素瘤M14细胞株的bel-2蛋白表达,有效抑制癌细胞的生长,为黑素瘤的治疗提供了一种新的治疗策略.
목적:탐토응용RNAi기술하조bcl-2기인표체후대인흑소류Mi4세포주생장적영향.방법:채용인흑소류M14세포주,이용인공합성적bel-2파향소분자간우RNA(small interferenceRNA,siRNA)전염세포,MTT법측세포증식정황.결과:면역조화가견25례실험조세포파편균표체bcl-2,표체강도약양성,25례공백대조조화음성대조균표체bel-2,표체강도강양성,실험조세포여공백대조조화음성대조조상비bcl-2표체명현감저,차이유통계학의의(P<0.05),공백대조조화음성대조조bcl-2표체차이무통계학의의(P>0.05),MTT법측정현시:전염48h후,실험조7조세포평균흡광도OD치0.384±0.026,공백대조조7조세포평균흡광도OD치0.590±0.032,음성대조조7조세포평균흡광도OD 치0.541±0.034,실험조세포흡광도강저,여공백대조조급음성대조조상비차이유통계학의의(P<0.05),이공백대조조화음성대조조량조간차이무통계학의의(P>0.05),실험조세포증식억제솔34.91%,음성대조조세포증식억제솔8.30%,차이유통계학의의(P<0.05),현시실험세포증식감만.결론:Bel-2RNAi능현저강저인흑소류M14세포주적bel-2단백표체,유효억제암세포적생장,위흑소류적치료제공료일충신적치료책략.
Objective: To study the inhibitory effect of a double-strand small interfering RNA targeting bcl-2siRNA in human melanoma cells. Method: Human melanoma cell M14 was treated with bcl2siRNA, the protein level of bcl-2 in the cells was determined by immunity class ; cell proliferation was detected by MTT. Result: Bcl-2 proteinum was buffy particle, most lied in cell endochylema. All twenty-five experimental groups cell expressed bcl-2, expression intensity was weakly positive, twenty-five blank groups and control groups expressed bcl-2, expression intensity was hadropositive,bcl-2 expression in experimental group was significantly lower than that in blank group and control group. There was no significantly difference in bcl-2 expression between blank group and control group. There was no bcl-2 expression in negative control group which used PBS replaced antibody. 48 hours after transfection , average absorbance of experimental group was 0.384±0.026, which was significantly lower than that in blank group ( 0.590±0.032 ) and control group(0.541±0.034 ), There's significanct the difference between experimental group and blank group or control group ( P < 0.05 ) , this shew transfection cell proliferation step down. Conclusion:Bcl-2RNAi can effectively knock down the bcl-2 protein and inhibit the growth of human melanoma cells,which might be a new way for melanoma .