人乳头状瘤病毒与促癌物协同作用诱发人胚宫颈细胞恶性转化研究

目的：研究人乳头状瘤病毒（HPV）与促癌物协同作用在scid小鼠体内诱发人胚宫颈细胞的恶性转化。方法：包装制备含HPV16E6E7逆转录病毒,感染人胚宫颈细胞,按分组将所需细胞移植于scid小鼠右侧肩部皮下,共分4组：实验组为感染病毒的宫颈细胞 亚精胺 正丁酸组,共7只;病毒组为感染病毒的宫颈细胞组,共5只;促癌组为正常宫颈细胞 亚精胺 正丁酸组,共5只;对照组为正常宫颈细胞组,共4只。按实验要求于移植第3日起在小鼠左侧肩部皮下注射亚精胺和正丁酸,每周1次。观察12周后处死动物,如发生肿瘤,则对瘤组织行病理诊断,并作PCR检测HPV16E6E7基因。结果：实验组成瘤比例5/7,其他3组成瘤率皆为0,实验组肿瘤的病理学检查证实为肉瘤,应用PCR方法在肿瘤组织中检测到HPV16E6E7基因。结论：宫颈细胞在感染含HPV16E6E7基因的逆转录病毒后,在亚精胺和正丁酸协同作用下发生恶性转化。

Induction of human cervical carcinoma by human papilloma virus and tumor promoters

Objective To study the effect of human papillomavirus (HPV) and tumour promoters spermidine (SPD and N butyrate) on malignant transformation of human embryo cervical cells. Methods Plasmid HPV16E6/E7 was constructed and transfected into human embryo cervical cells obtained by induced labor from pregnant women with severe heart diseases. Twenty one scid mice were divided into 4 groups: experimental group ( n =7, incubated subcutaneously with the transfected cervical cells at the right shoulder, and then injected subcutaneously with spermidine and N butyrate at the left shoulder three days after the incubation once a week for 12 times), virus infected group B ( n =5, incubated subcutaneously with transfected cervical cell only), tumor promoter group ( n =5, incubated with untransfected cervical cells and SPD and N butyrate), and control group ( n =4, incubated subcutaneously with only untransfected cervical cells). Twelve weeks after incubation, the mice were killed. Pathological rexamination was conducted to detect the existence of tumor. PCR was used to detect HPV16E6/E7 gene in tumor tissues. Result Tumor, diagnosed as fibrosarcoma by pathology, was found in 5 out of the 7 mice in the experimental group. The tumor forming rate was 0 in other groups. Expression of HPV16E6/E7 gene was detected in tumour tissues by PCR. Conclusion HPV16E6/E7 gene containing retrovirus infection, synergized by spermidine and N butyrate acid, causes malignant transformation in human cervical cells.