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首页> 外文期刊>European journal of cancer prevention: The official journal of the European Cancer Prevention Organisation (ECP) >Antisense human telomerase reverse transcriptase could partially reverse malignant phenotypes of gastric carcinoma cell line in vitro.
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Antisense human telomerase reverse transcriptase could partially reverse malignant phenotypes of gastric carcinoma cell line in vitro.

机译：反义人端粒酶逆转录酶可以部分逆转胃癌细胞系的恶性表型。

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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.
机译：在超过90％的检查肿瘤中检测到端粒酶活性，但在大多数正常体细胞中未检测到。在人类端粒酶的三个亚基中，人类端粒酶逆转录酶（hTERT）是端粒酶活性的限速成分。因此，靶向hTERT代表减少端粒酶功能的一种有前途的方法，它可能不会对端粒酶阴性的体细胞产生实质性的副作用。为了探索反义hTERT（ahTERT）对人SGC-7901胃癌细胞系恶性表型的影响，通过基因重组技术构建了hTERT反义真核表达载体。通过端粒重复扩增协议-ELISA检测端粒酶活性，通过逆转录PCR-检测端粒酶亚基，c-myc和bcl-2的mRNA，通过Southern印迹检测末端限制性片段（TRF），通过流式细胞术检测细胞周期分布和hTERT的蛋白表达转染前后，在SGC-7901细胞中通过蛋白质印迹法分析了c-myc和bcl-2。在上述细胞中，还检查并评估了软琼脂的克隆效率测定和裸鼠的肿瘤发生。结果表明，ahTERT转染后，SGC-7901细胞的增殖受到明显抑制。进一步的研究表明，在ahTERT转染的细胞中，端粒酶活性，端粒长度，hTERT，bcl-2和c-myc的mRNA和蛋白表达均降低。但是，在转染和未转染的细胞中，对人类端粒酶RNA（hTR）和人类端粒酶相关蛋白1（TP1）的转录均无明显影响。流式细胞仪分析显示在ahTERT转染的细胞中G0 / G1期积累，增殖指数（PI）下降。此外，在裸鼠中皮下注射ahTERT转染的细胞后未发现致瘤性，而在注射对照细胞的小鼠中观察到明显的肿瘤。我们的研究表明，外源的ahTERT可以通过抑制端粒酶活性，hTERT，c-myc和bcl-2表达来抑制SGC-7901细胞的增殖并部分逆转其恶性表型。针对hTERT策略的反义技术可能是胃癌治疗的潜在方法。

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