沉默HPV16 E6基因对人宫颈癌裸鼠移植瘤的抑制作用

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.

Inhibition effect of silencing HPV16 E6 gene on human cervical cancer CaSki cell carcinoma xenograft in nude mice

Objective To learn the effect on carcinoma xenograft in nude mice by inhibiting human papillomavirus 16(HPV16) E6 gene expression in CaSki cell. Methods The recombinant plasmids expressing HPV16 E6 small interference RNA (siRNA) were transfected into CaSki cell. The cells expressing recombinant plasmid was screened out with G418. The expression of E6 mRNA was determined by RT-PCR. The cells were inoculated into BALB/c nude mice subcutaneouly and the growth of the xenograft carcinoma was observed. After the pGensil-CH2 recombinant was injected into the carcinoma, the growth of carcinoma and pathological changes of carcinoma were observed. Results The CaSki cell expressing E6 siRNA was obtained, and HPV16 E6 mRNA expression in CaSki cell was down-regulated. The oncogenicity of the CaSki cell expressing E6 siRNA was degraded, the inhibition rate was up to 71.4% as compared with that of control group. The growth of tumor in nude mice was inhibited after the E6 siRNA plasmids were injected into the nude mice. The volume and weight of the tumor treated by siRNA were smaller than that of control group significantly. More necrotic area and less cell division phase were observed under light microscope in the E6 siRNA treated tumor. Conclusion The oncogenicity of the CaSki cell was degraded after silencing HPV16 E6 gene in CaSki cell by E6 siRNA.