[Establishment and characterization of a xenografted human cervical carcinoma in nude mice and homologous cell line in vitro]

Abstract

Objective: To establish a xenografted tumor strain of human cervical carcinoma in nude mice and homologous cell line in vitro.

Methods: A human well differentiated cervical squamous cell carcinoma derived from a surgical specimen was transplanted in the subcutis of nude mice. The xenograft was cultured to establish a cell line in vitro. A series of characteristics of the grafts and the cells were examined. The established cell line was cloned by single cell detaching flask. Non-serum culture of the cells was tried.

Results: The xenografts were successively passaged for 26 generations during a period of 17 months. A tumor strain HCC-94V was established, having a high rate of successful inoculation rate (92.9%) and was growing steadily. The morphological and biological characteristics of its original tumor were preserved. The chromosomal analysis revealed a human aneuploid pattern of 9-120 in number. The tumor markers detected by immunocytochemical techniques showed high expression, the oncogenic products, low expression. Polymerase chain reaction (PCR) showed the same results as those of the patients with original tumor tissue, human papilloma virus (HPV) 16 (+), 18 (-). A new human cell line HCC-94 derived from the xenograft, was established and maintained for 16 months through 131 passages. Transmission electromicroscopy demonstrated the presence of typical desmosomes between the cells and many tonofilaments in the cells. Also, the morphology of chromosomes showed the characteristics of chromosomes as well as of mutants in the human tumor cells. The tumor markers and the oncogenic products of the cells exhibited higher expression than that of the original tumor. The HPV 16 and 18 of the cells, using PCR, were negative. The cells were highly tumorigenic in nude mice. The transplanted tumors resembled the original tumor tissues in morphology and were HPV 16 positive. Both cell cloning culture and non-serum culture of HCC-94 were successful.

Conclusion: HCC-94V and HCC-94 could be considered as a new xenografted tumor strain may provide material and model ideal for further investigation of human cervical carcinoma.