Experimental Study On The Effect Of HTERT Promoter Regulated Vecter Targeting TRALL In Cervical Cancer Cells

The human telomerase reverse transcriptase (hTERT) has the effect of tumor-targeting.And Tumor necrosis factor (TNF) related apoptosis-inducing ligand (TRAIL) has been shown to induce apoptosis in a wide variety of cancer cells in vitro and to specifically limit tumor growth without damaging normal cells and tissues in vivo.These results suggest a strong potential of TRAIL as an anticancer therapy. Our research aimed to unclear whether a combination of hTERTpromoter regulated veter targeting TRALL and chemo-radiotherapy could enhance antitumor activity or could be used for the treatment of cervical cancer.AIMThe cervical cancer is one of the most common tumors of women. The aim of this research is to find the relationship of TRAILgene mutation with clinical metastasis of the cervical cancer; to find its effects on the mRNA or protein expression of TRAIL, cell proliferation, cell cycle, cell apoptosis, cell motion and invasion, and cell nodulation in the cervical cancer cell lines; to understand the mechanism between hTERT promoter and clinical metastasis of the cervical cancer; and to provide guidence to the therapy and prognosis of the cervical cancer.METHODSThe expression of TRAIL was examined in 10 cases of normal cervical tissue and 49 cases of the cervical carcinomas respectively by immunohistochemical technique; The expressions and methylation were classified and analyzed according to pathological grading, lymphatic metastasis and clinical staging. The mRNA or protein expression of TRAIL on HeLa and SiHa cell line was examined by RT-PCR and immunohistochemical staining respectively. The proliferation, cell cycle, apoptosis, motion, and invasion of the transfected cervical cancer cells were detected by MTT, flow cytometry, and cell invasion assay, respectively. Finally, the effects of TRAIL to the nodulation of the cervical cancer cell lines were detected by subcutaneous seeding in mice.RESULDSIn comparison with the normal cervical tissue, the expression of TRAIL was significantly decreased in the poorly differentiatied tumor and in the advanced patients (P<0.05), and also in the tumor tissues from the patients with local lymphnode metastases (P<0.05). On the contrary, the expression of TRAIL was significantly increased in all kinds of CIN(P<0.05), and it was downregulated along with the progressing of clinical stage (P<0.05); it was the same in the tumor tissues from the patients with local lymphnode metastases (P<0.05), but no differences were found in relation to cell differentiation (P>0.05). In the cervical cancer cell lines HeLa and SiHa, it could upregulate the G0/G1 stage percentage in cell cycle and could facilitate the apoptosis (P<0.05), and also inhibit the cell malignant proliferation, movement and invasion (P<0.05). Finally, Combined chemo-radiotherapy, hTERT-TRAIL could inhibit the growth of tumor nodulation by subcutaneous seeding in mice (P<0.05).CONCLUSIONThere was a close relationship between the TRAIL's upregulation with its malignant biological behavior; The TRAIL could significantly inhibit the malignant proliferation and invasion ability of the cervical cancer cell lines, facilitate the apoptosis and inhibit the tumor malignant proceeding. These results showed that hTERT-TRAIL had anti-tumor effect and could make an good foundation for the effective therapy of cervical cancer in the future. The use of hTERT-TRAIL to enhance sensitivity of cervical cancers to therapy represents an appealing therapeutic strategy worthy of further investigation.