SUBSCRIBE
Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2004, Vol. 2 ›› Issue (1): 42-45.doi: 10.3736/jcim20040116

• Original Experimental Research • Previous Articles     Next Articles

Effect of recombinant adenoviruses with CD/TK fusion suicide gene on human hepatocellular carcinoma cells

Chang-jun Wang1, Jian-jun Li2, Xiao-min Wen3, Li Deng1   

  1. 1. Department of Oncology, Guangzhou Hospital of Traditional Chinese Medicine, Guangzhou, Guangdong Province 510130, China
    2. Department of Integrated Traditional Chinese and Western Medicine, Second Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province 510120, China
    3. Department of Traditional Chinese Medicine, First Military Medical University, Guangzhou, Guangdong Province 510515, China
  • Online:2004-01-20 Published:2018-10-25

RichHTML

1

PDF (PC)

71

Cited

8

Like

Objective: To investigate gene-therapy for human hepatocellular carcinoma with adenovirus vectors by double suicide gene CD/TK. Methods: Double suicide gene CD/TK was liberated from eukaryotic vectors pCEA-CD/TK and subcloned into shuttle vectors, and the transfer plasmid pAdtrack-CMV-CD/TK was formed after linearizing with Pac 1. It was recombinated with pAdeasy-1 in bacteria BJ5183. The identified adenovirus plasmid was digested by Pac1 and was transfected into 293 cells to pack the adenoviruses. After PCR determination, its titre was measured, and the infection rate and efficacy were tested in human hepatocellular carcinoma cells. Results: pAdtrack-CMV-CD/TK and pAd-CD/TK were tested by endonuclease digestion. Ad-CD/TK was produced in 293 cells, and the human hepatocellular carcinoma cells (SMMC7721) infected by Ad-CD/TK were killed after 5-FC was used, and bystander effects were observed.Conclusion: Recombinant adenoviruses with CD/TK fusion suicide gene have a high infection rate and efficacy for human hepatocellular carcinoma cells.

Key words: adenovirus vector, gene therapy, double suicide gene, carcinoma, hepatocellular

CLC Number: 

  • R735.7

Fig.1

Electrophoresis identification of shuttleplasmid pAdtrack-CMV-CD/TK1:pCEA-CD/TK;2:pCEA-CD/TK HindⅢ/EcoRⅠdouble digestion （including 2.5 kb fragment of aimed genes and 5.4 kb fragment of carrier）;3:pCEA-CD/TK HindⅢ/BamHⅠdouble digestion（including about 1.3 kb fragment of CD and 6.6 kb fragment）;4:pAdtrack-CMV-CD/TK BamHⅠdigestion（12 kb）;5:pAdtrack-CMV-CD/TK HindⅢ/BamHⅠdouble digestion（6 kb,5 kb,1.3 kb）;M:DNA marker"

Fig.2

Electrophoresis identification of rAdplasmid pAd-CD/TK1:pAdtrack-CMV-CD/TK HindⅢ/BamHⅠdouble digestion（6 kb,5 kb,1.3 kb）;2:pAd-CD/TK PacⅠdigestion （30 kb and 4.5 kb）;3:pAdeasy-1 PacⅠdigestion（33 kb）;4:pAd-CD/TK HindⅢ/BamHⅠdouble digestion（including 1.3 kb fragment of CD）;M:DNA marker"

Fig.3

PCR identification of rAd plasmid Ad-CD/TK1:negative control;2:rAd （1.3 kb）;M:DNA marker"

Fig.4

Fluorescence photo of 293 cells afterAd-CD/TK infection"

[1] Freytag SO, Rogulski KR, Paielli DL , et al. A novel three-pronged approach to kill cancer cells selectively: concomitant viral, double suicide gene, and radiotherapy[J]. Hum Gene Ther, 1998,9(9):1323-1333
[2] Uckert W, Kammertons T, Haack K , et al. Double suicide gene(cytosine deaminase and herpes simplex virus thymidine kinase) but not single gene transfer allows reliable elimination of tumor cells in vivo[J]. Hum Gene Ther, 1998,9(6):855-865
[3] 许道松, 伍新尧, 钟女奇 , 等. 用融合自杀基因“靶向治疗”CEA阳性肿瘤[J]. 中山医科大学学报, 1998,19(1):79-80
[4] He TC , Zhou S, da Costa LT, et al.A simplified system for generating recombinant adenoviruses[J]. Proc Natl Acad Sci USA, 1998,95(5):2509-2514
[5] Danthinne X, Werth E . New tools for the generation of E1- and/or E3-substituted adenoviral vectors[J]. Gene Ther, 2000,7(1):80-87
doi: 10.1038/sj.gt.3301047 pmid: 10680020
[6] Danthinne X . New vectors for the construction of double recombinant adenoviruses[J]. J Virol Methods, 1999,81(1-2):11-20
doi: 10.1016/S0166-0934(99)00054-3 pmid: 10488756
[7] Graham FL, Prevec L . Methods for construction of adenovirus vectors[J]. Mol Biotechnol, 1995,3(3):207-220
[1] Gai Ran, Xi-lin Feng, Yi-lin Xie, Qing-yun Zheng, Peng-peng Guo, Ming Yang, Ying-lu Feng, Chen Ling, Li-qing Zhu, Chen Zhong. The use of miR122 and its target sequence in adeno-associated virus-mediated trichosanthin gene therapy. Journal of Integrative Medicine, 2021, 19(6): 515-525.
[2] Jia-jia Li, Qing Liang, Guang-chun Sun. Traditional Chinese medicine for prevention and treatment of hepatocellular carcinoma: A focus on epithelial-mesenchymal transition. Journal of Integrative Medicine, 2021, 19(6): 469-477.
[3] Tong Zhao, Bing-jie Guo, Chu-lan Xiao, Jiao-jiao Chen, Can Lü, Fan-fu Fang, Bai Li. Aerobic exercise suppresses hepatocellular carcinoma by downregulating dynamin-related protein 1 through PI3K/AKT pathway. Journal of Integrative Medicine, 2021, 19(5): 418-427.
[4] He-tong Zhao, Yong-bin Meng, Xiao-feng Zhai, Bin-bin Cheng, Sha-sha Yu, Man Yao, Hui-xia Yin, Xu-ying Wan, Yun-ke Yang, Hui Liu, Feng Shen, Chang-quan Ling. Comparable effects of Jiedu granules and sorafenib for advanced hepatocellular carcinoma: A prospective multicenter cohort study. Journal of Integrative Medicine, 2020, 18(4): 319-325.
[5] Porwornwisit Tritripmongkol, Tullayakorn Plengsuriyakarn, Mayuri Tarasuk, Kesara Na-Bangchang. In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component, ethyl-p-methoxycinnamate, against cholangiocarcinoma. Journal of Integrative Medicine, 2020, 18(4): 326-333.
[6] Kai-yue Tang, Shi-lin Du, Qian-lei Wang, Yi-fan Zhang, Hai-yan Song. Traditional chinese medicine targeting cancer stem cells as an alternative treatment for hepatocellular carcinoma. Journal of Integrative Medicine, 2020, 18(3): 196-202.
[7] Jeremiah Olorunjuwon Olugbami, Robert Damoiseaux, Bryan France, Michael A. Gbadegesin, Adam Z. Stieg, Shivani Sharma, Oyeronke A. Odunola, James K. Gimzewski. Atomic force microscopy correlates antimetastatic potentials of HepG2 cell line with its redox/energy status: Effects of curcumin and Khaya senegalensis. Journal of Integrative Medicine, 2017, 15(3): 214-230.
[8] Xuanbin Wang, Ning Wang, Fan Cheung, Lixing Lao, Charlie Li, Yibin Feng. Chinese medicines for prevention and treatment of human hepatocellular carcinoma: Current progress on pharmacological actions and mechanisms. Journal of Integrative Medicine, 2015, 13(3): 142-164.
[9] Yuan-hui Zhang, Yuan Wang, Yusufali Ali Hussein, Frederick Ashby, Daniel Zhang, Zi-fei Yin, George V. Aslanidi, Arun Srivastava, Chang-quan Ling, Chen Ling. Cytotoxic genes from traditional Chinese medicine inhibit tumor growth both in vitro and in vivo. Journal of Integrative Medicine, 2014, 12(6): 483-494.
[10] Zhi-yi Zhou, Ling Xu, He-gen Li, Jian-hui Tian, Li-jing Jiao, Sheng-fu You, Zhi-fen Han, Yi Jiang, Hui-ru Guo​, Hui Liu​. Chemotherapy in conjunction with traditional Chinese medicine for survival of elderly patients with advanced non-small-cell lung cancer: Protocol for a randomized double-blind controlled trial. Journal of Integrative Medicine, 2014, 12(3): 175-181.
[11] Chang-quan Ling​, Li-na Wang, Yuan Wang​, Yuan-hui Zhang​, Zi-fei Yin, Meng Wang​, Chen Ling​. The roles of traditional Chinese medicine in gene therapy. Journal of Integrative Medicine, 2014, 12(2): 67-75.
[12] Wang Li-na, Wang Yuan, Lu Yuan, Yin Zi-fei, Zhang Yuan-hui, V. Aslanidi George, Srivastava Arun, Ling Chang-quan, Ling Chen. Pristimerin enhances recombinant adeno-associated virus vector-mediated transgene expression in human cell lines in vitro and murine hepatocytes in vivo. Journal of Integrative Medicine, 2014, 12(1): 20-34.
[13] Hai-yan Song, Li Zhang, Jie-lu Pan, Li-li Yang, Guang Ji. Bioactivity of five components of Chinese herbal formula Jiangzhi granules against hepatocellular steatosis. Journal of Chinese Integrative Medicine, 2013, 11(4): 262-268.
[14] Xiao-feng Zhai, Zhe Chen, Bai Li, Feng Shen, Jia Fan, Wei-ping Zhou, Yun-ke Yang, Jing Xu, Xiao Qin, Le-qun Li, Chang-quan Ling. Traditional herbal medicine in preventing recurrence after resection of small hepatocellular carcinoma: A multicenter randomized controlled trial. Journal of Chinese Integrative Medicine, 2013, 11(2): 90-100.
[15] Shui-qiu Deng, Xue-nong Ouyang, Zong-yang Yu, Xi-hu Dai, Xi Chen, Fang-zheng Fang, Wen-wu Wang, Zhi-zhen Liu. Influence of Chinese herbal medicine Feitai Capsule on completion or delay of chemotherapy in patients with stage ⅢB/Ⅳ non-small-cell lung cancer: a Randomized controlled trial. Journal of Chinese Integrative Medicine, 2012, 10(6): 635-640.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. An explanation on "guiding principles of clinical research on mild cognitive impairment (protocol)". Journal of Chinese Integrative Medicine, 2008, 6(1): 15-21
[2] Yi-ting He, Qing-lin Zha, Jian-ping Yu, Yong Tan, Cheng Lu, Ai-ping Lv. Principal factor analysis of symptoms of rheumatoid arthritis and their correlations with efficacy of traditional Chinese medicine and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 32-36
[3] Jun Cai, Hua Wang, Sheng Zhou, Bin Wu, Hua-rong Song, Zheng-rong Xuan. Effect of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2008, 6(1): 37-40
[4] Wei Zhang, Xiang-feng Lu, Xiao-mei Zhang, Jian-jun Wu, Liang-duo Jiang. A rat model of pulmonary fibrosis induced by infusing bleomycin quickly through tracheal intubation. Journal of Chinese Integrative Medicine, 2008, 6(1): 60-67
[5] A-gao Zhou, Yong Zhang, Gang Kui, De-Yun Kong, Hai-liang Ge, Qiu-hua Ren, Jia-rong Dong, Sheng Hong, Xu-ming Mao, Yin Wang, Hui-zheng Zhang, Shu-jun Wang. Influence of traditional Chinese compound recipes with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 77-82
[6] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[7] Li Zhou, Hong-xing Zhang, Ling-guang Liu, Wen-jun Wan. Effect of electro-acupuncture at Fenglong (GV 16) on nitric oxide and endothelin in rats with hyperlipidemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 89-92
[8] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. Guiding principles of clinical research on mild cognitive impairment (protocol). Journal of Chinese Integrative Medicine, 2008, 6(1): 9-14
[9] Xin-jun Wang, Ling-ling Wang . A mechanism of endogenous opioid peptides for rapid onset of acupuncture effect in treatment of depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1014-1017
[10] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022
Journal of Integrative Medicine, ISSN: 2095-4964 CN 31-2083/R
Copyright © Journal of Integrative Medicine
Powered by Beijing Magtech Co. Ltd
沪公网安备31010602003416号      沪ICP备18017633号-3