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자궁경부암 세포주들과 Nude mouse 에 이종이식한 자궁경부암 세포에서 아데노바이러스와 리포펙틴을 이용한 종양억제 유전자 ( p53 ) 의 이입율
여경아(Kyeong A Yeo),정재은(Jae Eun Jung),이혜영(Hye Young Lee),이방현(Bang Hyon Lee),김태형(Tae Hyung Kim),천병수(Byeung Soo Cheung),노민석(Min Suk Rho),이준모(Joon Mo Lee),남궁성은(Sung Eun Namkoong),박용석(Yong Seok Park),김종국(Ch 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.6
N/A Background: The classical treatment of the cervical cancer is surgery, radiotherapy, chemotherapy. Even though the improvement of treatment successful rate, conventional therapy has some limitations. Recent cutting edge of cancer therapy has been developed in gene level including understand the biological characteristics of the cancer cells, enhance the human immune response, suppress the cancer cell proliferation. Therefore, the gene therapy is proposed to new treatment strategy. Purpose: The transfection efficiency of cervical cancer cell lines and cervical cancer cell line xerografted nude mouse was investigated by transfection of liposome and infection of adenovirus mediated suppressor(p53) and reportor(LacZ) gene. Method: The cervical cancer cell lines was used in this study were CaSki, SiHa (HPV16 positive, wild type p53 gene), HeLa, He]aS3(HPV18 positive, wild type p53 gene) and C33A, HT3(HPV negative, mutant p53). Direct plasmide and AdCMVp53 gene transfection was performed by using liposome system (pRcCMVLacZ / lipofectin, FuGene 6, Ca-phosphate). LacZ gene was used as the reportor gene for the transfection efficiency evaluation. Expression of p53 in cell lines and tumor tissue was confirmed by western blot and immunohistochemical staining. Xenografted nude mouse of SiHa cell line was infected by AdCMVp53 and AdCMVLacZ. Transfection efficiency was observed by same as above. Results: In cervical cancer cell lines, gene transfection using liposome system(pRcCMVLacZ/lipofectin, FuGene 6, Ca-phosphate)revealed different transfection efficiency, especially pRcCMVLacZ in Fugene 6 showed 18-40% of high transfection efficiency in 6 cervical cancer cell lines by X-gal staining and AdCMVp53 showed 95-98% of the high transfection efficiency in HeLa, C33A. AdCMVp53 was significantly expressed at 2-5days after injection xenografted nude mouse on the western blot and transfection efficiency was 19.79±5.36, 26.26 ± 11.69, 14.77±3.98,15.99 ±6.43%(day1-5). AdCMVLacZ were found to immunohistochemistry analysis, in vivo transFection efficiency was 61.26±4.66, 59.63±9.12, 29.46±14.33, 31.73±22.64%(day 1-5) at ×200 and 88.68.65, 70.85±20.94, 40.75±25.44, 48.21±10.97% (day 1-5) at ×400. Conclusion: As a results , adenovirus-mediated transfection efficiency was higher in vivo experiment compared to cell lines. These high efficiency of adenovirus-mediated suppressor gene(p53) could become a significant meaningful data gene therapy strategy both transgenic mice and cervical cancer cell lines.