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Inhibition of telomerase with human telomerase reverse transcriptase antisense increases the sensitivity of tumor necrosis factor-α-induced apoptosis in prostate cancer cells

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Asian Journal of Andrology 2007年第5期9卷 697-704页

作者： Xiao-Dong Gao Yi-Rong Chen Department of Urology Lanzhou University Lanzhou 730000 China Department of Urology People＇s Hospital of Gansu Province Lanzhou 730000 China

Aim： To investigate the effect of inhibition of telomerase with human telomerase reverse transcriptase （hTERT） antisense on tumor necrosis factor-α （TNF-α-induced apoptosis in prostate cancer cells （PC3）. Methods： Antisense phosphorothioate oligodeoxynucleotide （AS PS-ODN） was synthesized and purified. Telomerase activity was measured using the telomeric repeat amplification protocol （TRAP） and polymerase chain reaction enzyme-linked immunoassay （PCR-ELISA）. hTERT mRNA was measured by reverse transcription PCR （RT-PCR） assay and gel-image system, hTERT protein was detected by immunochemistry and flow cytometry. Cell viability was detected by 3-（4, 5-dimethylthiazol-2-yl）-2,5-Diphenyltetrazolium （MTT） assay. Cell apoptosis was observed by morphological method and determined by flow cytometry. Results： The telomerase activity decreased with time after hTERT AS PS-ODN treatment. The levels of hTERT mRNA decreased with time after hTERT AS PS-ODN treatment, which appeared before the decline of the telomerase activity. The percentage of positive cells of hTERT protein declined with time after hTERT AS PS-ODN treatment, which appeared after the decline of hTERT mRNA. There was no difference in telomerase activity, hTERT mRNA and protein levels between hTERT sense phosphorothioate oligodeoxynucleotide （S PS-ODN） and the control group. The cell viability decreased with time after hTERT AS PS-ODN combined with TNF-α treatment. The percentage of apoptosis increased with time after hTERT AS PS-ODN combined with TNF-α treatment. There was no difference in cell viability and the percentage of apoptosis between hTERT S PS-ODN and the control group. Conclusion： hTERT AS PS-ODN can significantly inhibit telomerase activity by downregulating the hTERT mRNA and protein expression, and inhibition of telomerase with hTERT antisense can enhance TNF-α- induced apoptosis of PC3 cells.

关键词： human telomerase reverse transcriptase antisense phosphorothioate oligodeoxynucleotide telomerase prostate cancer cells tumor necrosis factor-α

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Effect of miR-296 on the Apoptosis of Androgen-independent prostate cancer cells

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Journal of Reproduction and Contraception 2009年第1期20卷 1-10页

作者： Pei CHENG Run-sheng LI Biao-yang LIN Wei-qun WANG Yu-hua LI Yan GUO Wei LI Institute for Systems Biomedicine Shanghai Jiao Tong University Shanghai 200240 China Shanghai Institute of Planned Parenthood Research Shanghai 200032 China Zhejiang California International NanoSystems Institute Hangzhou 310029 China

Objective To investigate the miR-296＇s function in prostate carcinoma（PCa） cells. Methods In order to profile the miRNA expression in LNCaP cells, the cultured cells were stimulated with androgen after 48-h starvation, miRNA microarray analysis and Q-RT-PCR assay were performed. To characterize the effects of miR296 on PCa cells, CL-1 and PC-3 cells were transfected with miR-296 and antisense-miR-296, cell growth and apoptosis were then analyzed. Results The miR-296-5p expression was up-regulated by 2.22 folds in the CL-1 cells, which do not express significantly androgen receptor, than in LNCaP cells. Knockdown of miR-296-5p induced apoptosis of CL-1 cells, but not LNCaP cells. However, knockdown of miR-296-5p inhibited the growth rate of LNCaP cells cultured in absence of androgen. Conclusion MiR-296-5p could be important for development of prostate cancer from androgen dependence to androgen independence.

关键词： prostate cancer cells miR-296-5p apoptosis

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Recognition of Individual prostate cancer cells by the Combination of a Protein a-helix-sensitive Fluorescent Probe and a Zinc Sensor

Recognition of Individual Prostate Cancer Cells by the Combi...

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第八届全国化学生物学学术会议

作者： Nan Jiang Chanli Yang Xiongwei Dong Dan Zhao Dan Zhang Changlin Liu Key Laboratory of Pesticide ＆ Chemical Biology Ministry of EducationSchool of ChemistryCentral China Normal University

＜正＞prostate cancer cells have been reported to have both downregulated expression of the membrane proteins including zinc transporters and caveolin-1（Cav-1）and decreased zinc ion levels relative to pro state normal *** our study,we found that the benzazole derivatives 5-amino-2-benzooxazol-2-yl-phenol（1 in this paper）,which was reported to emit strong fluorescence in BSA resulted from an excited-state

关键词： Recognition prostate cancer cells α-helix fluorescent probe ESIPT Zinc sensor

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Exogenous p27KIP1 expression induces anti-tumour effects and inhibits the EGFR/PI3K/Akt signalling pathway in PC3 cells

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Asian Journal of Andrology 2009年第6期11卷 669-677页

作者： Jun Chen Dan Xia Jin-Dan Luo Ping Wang Department of Urology The First Affiliated Hospital Medical College of ZheJiang University Hangzhou 310003 China

p27 is a cyclin-dependent kinase inhibitor that regulates the progression of cells from G1 to S phase of the cell cycle. Loss of p27 has been associated with disease progression and with an unfavourable outcome in prostate cancer. In this study, we investigated whether exogenous p27 expression in the human androgen-independent prostate cancer PC3 cell line had any effect on cell growth, and we studied the molecular mechanisms involved. p27 expression was restored in PC3 cells by plasmid delivery. Cell proliferation and apoptosis were assessed in PC3 cells transfected with p27. We also investigated the effects of p27 on the epidermal growth factor receptor （EGFR）/ phosphatidylinositol 3-kinase （PI3K）/Akt signalling pathway in PC3 cells. By restoring p27 expression in PC3 cells, we observed that p27 reduced proliferation and induced arrest in G0/G1 phase. Moreover, p27-transfected PC3 cells underwent apoptosis, as shown by flow cytometric analysis and western blotting analysis of Bcl-2, Bax, Bad, caspase-3 and poly（ADP-ribose）polymerase expression. Furthermore, the p27-induced anti-tumour action corre- lated with inhibition of the EGFR/PI3K/Akt signalling pathway, as confirmed by western blotting analysis and densitometry of EGFR, PI3K （p85）, Akt and p-Akts473 expression. Our results suggest that exogenous expression of p27 inhibits the proliferation of PC3 cells through induction of G1 arrest and apoptosis, and this process correlates with inhibition of the EGFR/PI3K/Akt signalling pathway.

关键词： apoptosis EGFR p27 prostate cancer cells signalling pathway

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