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Yang C.-R.,General Hospital | Wang Y.-D.,General Hospital | Jing S.-W.,Hebei Medical University | Gao J.-M.,General Hospital | And 2 more authors.
Tumor | Year: 2013

Objective: To investigate the effects of UHRF1 (ubiquitin-like with PHD and ring finger domains 1) gene silencing by shRNA (short hairpin RNA) interference on the biological behaviors of esophageal carcinoma TE-1 cells. Methods: The protein expression levels of UHRF1 in esophageal carcinoma tissues and normal esophageal tissues were detected by immunohistochemical staining assay to confirm overexpression of UHRF1 in esophageal carcinoma tissues. After UHRF1-shRNA was transfected into TE-1 cells by method of lentiviral infection, the mRNA and protein levels of UHRF1 in TE-1 cells were detected by RT-PCR (reverse transcription-PCR) and Western blotting, respectively. The proliferative and invasive capabilities of TE-1 cells transfected with UHRF1-shRNA were detected by the MTT assay and Transwell invasion assay, respectively. The cell cycle distribution and apoptosis were determined by FCM (flow cytometry). Results: UHRF1 was overexpressed in esophageal carcinoma tissues, as compared with the normal esophageal tissues (low or absent expression, P < 0.01). After transfection with UHRF1-shRNA, the mRNA and protein levels of UHRF1 in TE-1 cells were significantly reduced, the proliferative and invasive capabilities of TE-1 cells were inhibited, and the cell cycle was arrested in G0/G1 phase (P < 0.05) with an increase in apoptosis (P < 0.05). Conclusion: Small RNA interference targeting UHRF1 gene can inhibit the proliferation, reduce the invasive capability, and induce the apoptosis of TE-1 cells, indicating that UHRF1 plays an important role in oncogenesis and progression of human esophageal carcinoma. Copyright © 2013 by TUMOR.

Wang P.,Peoples Hospital of Tangshan | Wu X.-H.,Shijiazhuang University | Shan B.-E.,Shijiazhuang University
Chinese Journal of Cancer Prevention and Treatment | Year: 2011

OBJECTIVE: To detect the effects of lysophosphatidic acid (LPA) on MMP-2 and MMP-9 expression and activity in ovarian cancer cells with stable expression LPA2 and LPA3 genes. METHODS: pcDNA3. 1flag-LPA2 and pcDNA3. 1flag-LPA3 were transfected into ovarian cancer cells lines (SKOV3 and 3AO) with lipofectamine reagent. Transfection and expression were confirmed by the means of Western blotting. The effect of lysophosphatidic acid (LPA) on MMP-2 and MMP-9 protein expression and activity in transfected ovarian cells was analyzed by gelatin zymography. RESULTS: High expression of LPA2 and LPA3 protein was analyzed by Western blotting. The gelatin zymography confirmed that LPA- induced the activity and protein expression of MMP-2 and MMP-9 increased significantly in the transfected LPA2 and LPA3 receptors genes cells, compared with that in the control groups and vector groups (P<0.05). CONCLUSIONS: It is confirmed that ovarian carcer cells with high expression of LPA2 and LPA3 genes in vitro have been established. LPA2 and LPA3 are more efficient in LPA-induced MMPs activity. LPA2 and LPA3 play an important role in the process of LPA-induced MMP invasion and metastasis of ovarian carcer cells.

Yang C.,Hebei Medical University | Wang Y.,Hebei Medical University | Zhang F.,Military General Hospital of Beijing PLA | Sun G.,Peoples Hospital of Tangshan | And 4 more authors.
Molecular Biology Reports | Year: 2013

Radiotherapy is an effective treatment for some esophageal cancers, but the molecular mechanisms of radiosensitivity remain unknown. Ubiquitin-like with PHD and ring finger domains 1 (UHRF1) is a novel nuclear protein which is overexpressed in various cancers but not yet examined in esophageal squamous cell carcinoma (ESCC). The correlation between UHRF1 and the radioresistance in ESCC is still unclear. In the present study, the expression of UHRF1 was examined by immunohistochemistry in specimens of ESCC patients treated with radiotherapy. The results showed that UHRF1 was significantly overexpressed in ESCC specimens. Overexpression of UHRF1 correlated significantly with advanced T-stage, positive lymph node metastasis and poor differentiation. In addition, UHRF1 was associated with radiotherapy response, in which overexpression of UHRF1 was observed more frequently in the radioresistant group than in the effective group. At the molecular level, inhibition of UHRF1 by lentivirus-mediated shRNA targeting UHRF1 increased the radiosensitivity and apoptosis, while decreased radiation-induced G2/M phase arrest in TE-1 cells. Moreover, inhibition of UHRF1 resulted in higher residual γH2AX expression after irradiation, but not initial γH2AX. Further study showed that inhibition of UHRF1 down-regulated the endogenous expressions of DNA repair protein Ku70 and Ku80 in TE-1 cells, and significantly inhibited the increase of these proteins after irradiation. Above all, our data suggested that UHRF1 might play an important role in radioresistance of ESCC, and inhibition of UHRF1 can increase the radiosensitivity of TE-1 cells by altering cell cycle progression, enhancing apoptosis, and decreasing DNA damage repair capacity. © 2013 Springer Science+Business Media Dordrecht.

Li C.,Hebei Medical University | Zhou X.,Hebei Medical University | Wang Y.,Hebei Medical University | Jing S.,Hebei Medical University | And 5 more authors.
Molecular Medicine Reports | Year: 2014

micro (mi)RNAs are short regulatory RNAs that negatively modulate protein expression at the post-transcriptional level, and are being considered as novel therapeutic targets for the treatment of cancer. In the present study, an elevated expression level of circulating miR-210 was observed in patients with esophageal squamous cell carcinoma (ESCC) for the first time, to the best of our knowledge, and the induction of miR-210 under hypoxic conditions in ESCC was confirmed. Cell counting kit-8 assay and bromodeoxyuridine incorporation assay indicated that miR-210 markedly inhibited the proliferation of ESCC cells. In addition, the effect of miR-210 on the cell cycle was examined. Transfection of miR-210 resulted in a significant increase in the proportion of cells in G 2/M phase. Polo-like kinase 1 (PLK1) was investigated as a candidate target of miR-210, which is a critical regulator of cell cycle transmission at multiple levels. It was demonstrated that miR-210 reduced the levels of PLK1 protein by binding the 3′ untranslated region of its mRNA. The results of the present study demonstrated that miR-210 inhibited the proliferation of ESCC cells by inducing G2/M phase cell cycle arrest, and these effects of miR-210 were mediated by the targeting of PLK1.

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