机构:[1]Institute of Gastroenterology of PLA Southwest Hospital, Third Military Medical University, Chongqing[2]Institute of Burn Research of PLA, Southwest Hospital, Third Military Medical University, Chongqing[3]Key Laboratory of Biotherapy of Human Diseases, Ministry of Education[4]Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China四川大学华西医院
Telomerase activity is detected in more than 90% of examined tumors but not in most normal somatic cells. Among three subunits of human telomerase, human telomerase reverse transcriptase (hTERT) is the rate-limiting component for telomerase activity. Therefore, targeting hTERT represents a promising approach for diminishing telomerase function that will probably not cause substantial side effects on telomerase negative somatic cells. To explore the effects of antisense hTERT (ahTERT) on the malignant phenotypes of human SGC-7901 gastric cancer cell line in vitro, an antisense eukaryotic expression vector of hTERT was constructed by gene recombinant technology. Telomerase activity by telomeric repeat amplification protocol-ELISA, mRNA of telomerase subunits, c-myc and bcl-2 by reverse transcript-PCR, terminal restriction fragment (TRF) by Southern blot, cell cycle distribution by flow cytometry and protein expression of hTERT, c-myc and bcl-2 by Western blot were analyzed in SGC-7901 cells before and after transfection. Cloning efficiency assay in soft agar and tumorigenesis in nude mice were also examined and evaluated in the above cells. The results demonstrated that after ahTERT transfection, the proliferation of SGC-7901 cells was significantly inhibited. Further study showed that telomerase activity, telomere length, the mRNA and protein expression of hTERT, bcl-2 and c-myc were decreased in ahTERT-transfected cells. There were, however, no obvious effects on transcription of human telomerase RNA (hTR) and human telomerase associated protein1 (TP1) in both transfected and untransfected cells. Flow cytometric analysis displayed an accumulation of G0/G1 phase and a decreasing proliferation index (PI) in ahTERT-transfected cells. Moreover, no tumorigenicity was found after subcutaneous injection of ahTERT-transfected cells in nude mice, whereas palpable tumors were observed in mice injected with control cells. Our study indicates that exogenous ahTERT can inhibit proliferation and partially reverse malignant phenotypes of SGC-7901 cells via the suppression of telomerase activity, hTERT, c-myc and bcl-2 expression. Antisense technology targeted hTERT strategy might be a potential approach for gastric cancer therapy.
基金:
National Nature Science
Foundation of China grant 39770302, 30170425 and
30470797.
语种:
外文
PubmedID:
中科院(CAS)分区:
出版当年[2008]版:
大类|4 区医学
小类|4 区肿瘤学
最新[2023]版:
大类|4 区医学
小类|4 区肿瘤学
第一作者:
第一作者机构:[1]Institute of Gastroenterology of PLA Southwest Hospital, Third Military Medical University, Chongqing[*1]Institute of Gastroenterology of PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
通讯作者:
通讯机构:[1]Institute of Gastroenterology of PLA Southwest Hospital, Third Military Medical University, Chongqing[*1]Institute of Gastroenterology of PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
推荐引用方式(GB/T 7714):
Yang Shi Ming,Fang Dian Chun,Yang Jin Liang,et al.Antisense human telomerase reverse transcriptase could partially reverse malignant phenotypes of gastric carcinoma cell line in vitro.[J].European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation (ECP).2008,17(3):209-17.doi:10.1097/CEJ.0b013e3282b71f0d.
APA:
Yang Shi Ming,Fang Dian Chun,Yang Jin Liang,Chen Ling,Luo Yuan Hui&Liang Guang Ping.(2008).Antisense human telomerase reverse transcriptase could partially reverse malignant phenotypes of gastric carcinoma cell line in vitro..European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation (ECP),17,(3)
MLA:
Yang Shi Ming,et al."Antisense human telomerase reverse transcriptase could partially reverse malignant phenotypes of gastric carcinoma cell line in vitro.".European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation (ECP) 17..3(2008):209-17