BLTR-4永生化细胞系的建立及生物学特性初步鉴定

Establishment of Immortalized Cell Line BLTR-4 and PrimaryIdentification of Its Biological Character

目的建立膀胱尿路上皮永生化细胞系,并初步鉴定其生物学特性。方法以Fugene-6TM为载体,将含人乳头瘤病毒16(HPV-16雪E6、E7基因的HPV-16K质粒,体外转染原代培养的正常胎儿膀胱尿路上皮细胞。运用PCR、免疫组织化学及细胞特征鉴定等方法,对所得BLTR-4细胞系中HPV-16E6、E7基因的存在情况及该细胞系生物学特性进行初步鉴定。结果BLTR-4细胞系是含有HPV-16E6、E7基因并表达上述两种蛋白的尿路上皮源性细胞系。该细胞在体外已传70余代,细胞倍体数发生了二倍体至四倍体再至非整倍体的改变,生长特性符合永生化细胞系。结论BLTR-4细胞系是膀胱尿路上皮永生化细胞系,该细胞系含有HPV-16E6、E7基因。BLTR-4细胞系的建立为研究高危型HPV感染与膀胱移行细胞癌穴TCC雪关系提供了体外实验模型。

Objective To establish immortalized cell line from the urothelium of the urinary bladder and identify the characteristics of the cell line. Methods Human papillomavirus 16?(HPV-16?)plasmid was used to transfect urothelium of infant urinary bladder in vitro with the help of Fugene-6TM? and this plasmid contained E6 and E7 genes of HPV-16. We also identified the existence of HPV-16 E6 and E7 genes and the biological characteristics of the cell line by PCR? immunohistochemistry? and the biology identification. Results BLTR-4 cell line? produced from the transfection of HPV-16K plasmid? was a cell line from urothelium with the expression of HPV-16 E6 and E7 genes. It had been cultured more than 70 passages? and the characteristics of growth was similar to the immortalized cell line as reported. Conclusions BLTR-4 cell line is an immortalized cell line from urothelium of the urinary bladder? which contains HPV-16 E6 and E7 genes. BLTR-4 cell line is a good experimental model to investigate the relationship of the infection of high risk HPV and transitional cell carcinoma?(TCC?)in vitro.