CAJ | 학술논문

目的探讨RNA激活技术上调p21WAF1/CIP1基因的表达而影响人肝癌细胞株BEL-7402的增殖和凋亡能力.方法将与p21WAF1/CIP1基因启动子区域DNA序列互补的RNA分子(dsP21-322)转染入BEL-7402细胞中,采用qPCR法及Western印迹方法检测p21 WAF1/CIP1的表达;采用WST-1细胞增殖试剂检测细胞增殖情况;使用流式细胞仪评估肿瘤细胞凋亡情况并使用Western印迹方法检测相关分子表达水平的变化.结果 dsP21-322转染72 h后,BEL-7402细胞中p21 WAF1/CIP1表达显著上调;细胞增殖受到明显抑制,凋亡率36.86％(细胞的体积变小、变形,细胞核固缩,染色质凝聚呈深染,细胞出现皱缩、变圆、脱落)较对照组(11.51％、14.06％)相比明显升高.结论 RNA激活技术靶向上调p21WAF1/CIP11基因表达并抑制细胞增殖、增加诱导肿瘤细胞凋亡,可作为肝细胞癌或其他恶性肿瘤基因治疗的一种有效手段.
목적 탐토RNA격활기술상조p21WAF1/CIP1기인적표체이영향인간암세포주BEL-7402적증식화조망능력.방법 장여p21WAF1/CIP1기인계동자구역DNA서렬호보적RNA분자(dsP21-322)전염입BEL-7402세포중,채용qPCR법급Western인적방법검측p21 WAF1/CIP1적표체;채용WST-1세포증식시제검측세포증식정황;사용류식세포의평고종류세포조망정황병사용Western인적방법검측상관분자표체수평적변화.결과 dsP21-322전염72 h후,BEL-7402세포중p21 WAF1/CIP1표체현저상조;세포증식수도명현억제,조망솔36.86％(세포적체적변소、변형,세포핵고축,염색질응취정심염,세포출현추축、변원、탈락)교대조조(11.51％、14.06％)상비명현승고.결론 RNA격활기술파향상조p21WAF1/CIP11기인표체병억제세포증식、증가유도종류세포조망,가작위간세포암혹기타악성종류기인치료적일충유효수단.
Objective To evaluate the anti neoplastic effects of p21WAF1/CIP1 transcriptional activation induced by duplex RNAs in hepatocellular carcinoma (HCC) cell line BEL-7402.Methods Cells were treated with dsRNAs complementary to promoter sequences of p21WAF1/CIP1.Quantitative polymerase chain reaction (qPCR) and Western blot were employed to detect the expression of p21.At various timepoints post-transfection,cell viability assay and apoptosis analysis were used to determine the effect of RNA activation.After transfection Western blot was also performed to detect the expression of BclxL,cleaved caspase-3,cleaved caspase-9 and cleaved PARP.Results DsP21-322 transfection significantly inhibited cell viability.And,at Day 5,dsP21-322 inhibited cell growth by 65.84％ versus control.Flow cytometry revealed that dsP21-322 caused a significant increase of cell apoptosis.The total percent of apoptotic cells (UR + LR) increased to 36.86％ versus 11.51％ and 14.06％ in mocks and controls respectively.Such phenomena correlated with a decrease of anti-apoptotic protein Bcl-xL and an increase of cleaved caspase-3,cleaved caspase-9 and cleaved PARP.Conclusion Activation of p21 gene expression by saRNA may offer therapeutic benefits for HCC and other cancers.