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        Genotype­specific methylation of HPV in cervical intraepithelial neoplasia

        Yaw-Wen Hsu,Rui-Lan Huang,Po-Hsuan Su,Yu-Chih Chen,Hui-Chen Wang,Chi-Chun Liao,Hung-Cheng Lai 대한부인종양학회 2017 Journal of Gynecologic Oncology Vol.28 No.4

        Objective: Hypermethylation of human papillomavirus (HPV) and host genes has beenreported in cervical cancer. However, the degree of methylation of different HPV typesrelative to the severity of the cervical lesions remains controversial. Studies of the degree ofmethylation associated with the host gene and the HPV genome to the severity of cervicallesions are rare. We examined the association of methylation status between host genes andlate gene 1 (L1) regions of HPV16, 18, 52, and 58 in cervical brushings. Methods: Cervical brushings from 147 HPV-infected patients were obtained. The samplescomprised normal (n=28), cervical intraepithelial neoplasia (CIN) 1 (n=45), CIN2 (n=13), andCIN3/carcinoma in situ (n=61). The methylation status of HPV and host genes was measuredusing bisulfite pyrosequencing and quantitative methylation-specific polymerase chainreaction (PCR). Results: The degree of methylation of L1 in HPV16, 18, and 52 was associated with theseverity of the cervical lesion. In HPV52, C-phosphate-G (CpG) sites 6368m, 6405m, and6443m showed significantly higher methylation in lesions ≥CIN3 (p=0.005, 0.003, and0.026, respectively). Methylation of most HPV types except HPV52 (r<−0.1) was positivelycorrelated with the degree of methylation of host genes including PAX1 and SOX1 (0.4≤r≤0.7). Combining HPV methylation with PAX1 methylation improved the clustering for ≥CIN2. Conclusion: Our study showed that the degree of L1 methylation of HPV16, 18, and 52but not 58 is associated with the severity of cervical lesions. The association betweenHPV methylation and host gene methylation suggests different responses of host cellularepigenetic machinery to different HPV genotypes.

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        Vimentin Is Involved in Peptidylarginine Deiminase 2-Induced Apoptosis of Activated Jurkat Cells

        Hsu, Pei-Chen,Liao, Ya-Fan,Lin, Chin-Li,Lin, Wen-Hao,Liu, Guang-Yaw,Hung, Hui-Chih Korean Society for Molecular and Cellular Biology 2014 Molecules and cells Vol.37 No.5

        Peptidylarginine deiminase type 2 (PADI2) deiminates (or citrullinates) arginine residues in protein to citrulline residues in a $Ca^{2+}$-dependent manner, and is found in lymphocytes and macrophages. Vimentin is an intermediate filament protein and a well-known substrate of PADI2. Citrullinated vimentin is found in ionomycin-induced macrophage apoptosis. Citrullinated vimentin is the target of anti-Sa antibodies, which are specific to rheumatoid arthritis, and play a critical role in the pathogenesis of the disease. To investigate the role of PADI2 in apoptosis, we generated a Jurkat cell line that overexpressed the PADI2 transgene from a tetracycline-inducible promoter, and used a combination of 12-O-tetradecanoylphorbol-13-acetate and ionomycin to activate Jurkat cells. We found that PADI2 overexpression reduced the cell viability of activated Jurkat cells in1a dose- and time-dependent manner. The PADI2-overexpressed and -activated Jurkat cells presented typical manifestations of apoptosis, and exhibited greater levels of citrullinated proteins, including citrullinated vimentin. Vimentin overexpression rescued a portion of the cells from apoptosis. In conclusion, PADI2 overexpression induces apoptosis in activated Jurkat cells. Vimentin is involved in PADI2-induced apoptosis. Moreover, PADI2-overexpressed Jurkat cells secreted greater levels of vimentin after activation, and expressed more vimentin on their cell surfaces when undergoing apoptosis. Through artificially highlighting PADI2 and vimentin, we demonstrated that PADI2 and vimentin participate in the apoptotic mechanisms of activated T lymphocytes. The secretion and surface expression of vimentin are possible ways of autoantigen presentation to the immune system.

      • KCI등재

        Vimentin Is Involved in Peptidylarginine Deiminase 2-Induced Apoptosis of Activated Jurkat Cells

        Pei-Chen Hsu,Ya-Fan Liao,Chin-Li Lin,Wen-Hao Lin,Guang-Yaw Liu,Hui-Chih Hung 한국분자세포생물학회 2014 Molecules and cells Vol.37 No.5

        Peptidylarginine deiminase type 2 (PADI2) deiminates (or citrullinates) arginine residues in protein to citrulline residues in a Ca2+-dependent manner, and is found in lymphocytes and macrophages. Vimentin is an intermediate filament protein and a well-known substrate of PADI2. Citrullinated vimentin is found in ionomycin-induced macrophage apoptosis. Citrullinated vimentin is the target of anti-Sa antibodies, which are specific to rheumatoid arthritis, and play a critical role in the pathogenesis of the dis-ease. To investigate the role of PADI2 in apoptosis, we generated a Jurkat cell line that overexpressed the PADI2 transgene from a tetracycline-inducible promoter, and used a combination of 12-O-tetradecanoylphorbol-13-acetate and ionomycin to activate Jurkat cells. We found that PADI2 overexpression reduced the cell viability of activated Jurkat cells in a dose- and time-dependent manner. The PADI2-overexpressed and -activated Jurkat cells presented typical manifestations of apoptosis, and exhibited greater levels of citrullinated proteins, including citrullinated vimentin. Vimentin overexpression rescued a portion of the cells from apoptosis. In conclusion, PADI2 overexpression induces apoptosis in activated Jurkat cells. Vimentin is involved in PADI2-induced apoptosis. Moreover, PADI2-overexpressed Jurkat cells secreted greater levels of vimentin after activation, and expressed more vimentin on their cell surfaces when undergoing apoptosis. Through artificially highlighting PADI2 and vimentin, we demonstrated that PADI2 and vimentin participate in the apoptotic mechanisms of activated T lymphocytes. The secretion and surface expression of vimentin are possible ways of autoantigen presentation to the immune system.

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