The study was performed to construct a human cervical cancer cell line C33A which can stably express HPV58E6E7 fusion gene. Firstly, C33A cells were transfected with the recombinant lentivirus LV-HPV58E6E7 which contained HPV58E6E7 fusion gene, and the stably transfected cells (LV-HPV58E6E7/C33A) were screened out by flow cytometry. MTT was used to observe the growth of LV-HPV58E6E7/C33A cells and flow cytometry was carried out to detect the cell cycle. LV-HPV58E6E7/C33A cells were inoculated into the left armpits of nude mice. Then, the transcription and expression of HPV58E6E7 fusion gene was detected by qRT-PCR and Western blot, respectively. The results showed that HPV58E6E7 fusion gene can promote the proliferation of C33A cells. HPV58E6E7 fusion gene can be stably transcripted and expressed in vaccinated nude mice. The conclusion indicated that we successfully established a cervical cancer cell line LV-HPV58E6E7/C33A which can stably express HPV58E6E7 fusion gene. This cell line will provide an antigen cell line for the immune effect detection of HPV58 therapeutic vaccine.
Citation：
JIA Chaoying, MA Weihong, WANG He. Establishment and identification of human cervical cancer cell line C33A stably expressing human papillomavirus type 58 E6E7 fusion gene. Journal of Biomedical Engineering, 2017, 34(4): 578-584. doi: 10.7507/1001-5515.201605040
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Copyright © the editorial department of Journal of Biomedical Engineering of West China Medical Publisher. All rights reserved
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杨秀坤, 陈宣男, 姜铮. pcDNA3 转染 HPV16E6 基因对人宫颈癌细胞 C33A 细胞生长及周期的影响. 第三军医大学学报, 2009, 31(9): 835-838.
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- 1. 乔友林. 中国妇女人乳头瘤病毒感染和子宫颈癌的流行病学研究现状及其疫苗预防前景. 中华流行病学杂志, 2007, 28(10): 937-940.
- 2. McCloskey R, Menges C, Friedman A, et al. Human papillomavirus type 16 E6/E7 upregulation of nucleophosmin is important for proliferation and inhibition of differentiation. J Virol, 2010, 84(10): 5131-5139.
- 3. Lombard I, Vincent-Salomon A, Validire P, et al. Human papillomavirus genotype as a major determinant of the course of cervical Cancer. J Clin Oncol, 1998, 16(8): 2613-2619.
- 4. 赵瑾, 王小萼, 吕磊, 等. 不同宫颈病变中人乳头状瘤病毒的型别分布研究. 中华医院感染学杂志, 2011, 21(10): 2027-2033.
- 5. 马伟红, 贾超颖,王鹤. 表达人乳头瘤病毒 58 型(HPV58)E6E7 融合基因的慢病毒系统的建立及真核表达. 南宁: 广西医科大学, 2017.
- 6. 周慧, 李莉. IL-24 对人宫颈癌 C33A 细胞裸鼠移植瘤的治疗研究. 中国医疗前沿, 2011, 6(6): 45-46, 34.
- 7. Cornet S, Menez-Jamet J, Lemonnier F, et al. CpG oligodeoxynucleotides activate dendritic cells in vivo and induce a functional and protective vaccine immunity against a TERT derived modified cryptic MHC class I-restricted epitope. Vaccine, 2006, 24(11): 1880-1888.
- 8. 张艾芃. HPV16/18 E6/E7 基因沉默对 Siha/Hela 细胞生物学行为影响的研究. 北京: 军医进修医院, 2012.
- 9. 杨秀坤, 陈宣男, 姜铮. pcDNA3 转染 HPV16E6 基因对人宫颈癌细胞 C33A 细胞生长及周期的影响. 第三军医大学学报, 2009, 31(9): 835-838.
- 10. Byrne H M. Dissecting cancer through mathematics: from the cell to the animal model. Nat Rev Cancer, 2010, 10(3): 221-230.
