论文部分内容阅读
目的采用RNA干扰抑制人宫颈癌CaSki细胞裸鼠移植瘤中人乳头瘤病毒(human papillomavirus-16,HPV16)E6基因的表达,观察对裸鼠移植瘤的抑制作用。方法将HPV16 E6特异性的小干扰RNA(small interference RNA,siRNA)真核表达载体转染到人宫颈癌细胞株CaSki细胞中,经G418筛选,RT-PCR检测E6基因的表达;将细胞接种到BALB/c裸鼠背部皮下,同时将该质粒注射到人宫颈癌CaSki细胞移植瘤的裸鼠中,观察肿瘤出现时间及体积变化;观察肿瘤组织的病理改变。结果成功筛选到持续表达E6 siRNA的CaSki细胞,经RT-PCR检测该细胞中HPV16 E6基因表达降低;裸鼠成瘤实验结果显示,处理后的CaSki细胞成瘤能力明显降低,抑瘤率为71.4%;注射质粒进行基因治疗后,裸鼠肿瘤生长受到抑制,肿瘤的体积与质量明显低于CaSki细胞组,差异具有显著性(P<0.05)。病理检查结果显示,经pGensil-CH2处理的肿瘤生长不佳,细胞分裂像少见,组织内有较多坏死区域。结论沉默CaSki细胞中HPV16 E6基因的表达能明显降低宫癌细胞的成瘤作用。
OBJECTIVE: To study the inhibitory effect of RNA interference on E6 gene expression in human papillomavirus-16 (HPV16) xenografts in nude mice bearing cervical cancer. Methods Eukaryotic expression vector of HPV16 E6-specific small interfering RNA (siRNA) was transfected into human cervical cancer cell line CaSki. The cells were screened by G418 and the expression of E6 gene was detected by RT-PCR. The BALB / c nude mice were injected subcutaneously into the back of the nude mice. At the same time, the plasmid was injected into the nude mice bearing human cervical cancer CaSki cell transplanted tumor. The time and volume change of the tumor were observed. The pathological changes of the tumor were observed. Results CaSki cells with continuous expression of E6 siRNA were successfully screened and the expression of HPV16 E6 gene was decreased by RT-PCR. The tumorigenicity of CaSki cells in nude mice was significantly reduced, with the inhibition rate of 71.4 %. After gene therapy, the growth of tumor in nude mice was inhibited and the volume and quality of tumor were significantly lower than that of CaSki cells (P <0.05). The results of pathological examination showed that the tumors treated with pGensil-CH2 showed poor growth, rare cell division and more necrotic area in the tissue. Conclusion The silencing of HPV16 E6 gene expression in CaSki cells can significantly reduce the tumorigenicity of uterine cancer cells.