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Wang S.-S.,Guangdong Pharmaceutical University | Yang Q.-S.,Guangdong Pharmaceutical University | Shi J.-W.,Guangdong Pharmaceutical University | Shi J.-W.,Guangdong Province Key Laboratory for Biotechnology Drug Candidates | And 3 more authors.
Tumor | Year: 2014

Objective: To investigate the stable expression of voltage-gated chloride channel 3 (ClC-3) on the sensitivity to cisplatin in cervical cancer HeLa cells, and to explore its possible mechanism. Methods: The recombination vector plasmid pcDNA3.1/ClC-3 was transfected into HeLa cells by liposome-mediated intracellular delivery. After transfection with pcDNA3.1/ClC-3, the expression levels of ClC-3 protein and P-glycoprotein (P-gp) in HeLa cells were determined by Western blotting, and the expression levels of ClC-3 and multi-drug resistance gene 1 (MDR1) mRNAs were examined by real-time fluorescence quantitative PCR. MTT assay was used to determine the growth inhibition induced by cisplatin, and then the expressions and location of ClC-3 protein and P-gp were observed by immunofluorescence. Results: The ClC-3 was stably expressed in HeLa cells, which was named as HeLa/ClC-3. The expression levels of MDR1 and ClC-3 mRNAs as well as ClC-3 protein and P-gp in HeLa/ClC-3 cells were higher than those in HeLa/mock cells transfected with empty vector pcDNA3.1 (P < 0.05). As compared with the HeLa/mock cells, the inhibitory effect of cisplatin on HeLa/ClC-3 cells decreased obviously (P < 0.05), and the half inhibitory concentration (IC50) value of cisplatin in HeLa/ClC-3 cells was higher than that in HeLa/mock cells (71 μmol/L vs 32 μmol/L, P < 0.05). The ClC-3 protein and P-gp were co-located at the cell membrane of HeLa/ClC-3 and HeLa/mock cells. Conclusion: ClC-3 can reduce the sensitivity of HeLa cells to cisplatin. This effect is associated with up-regulation of P-gp expression. Copyright© 2014 by Tumor.

Shao H.,Guangdong Province Key Laboratory for Biotechnology Drug Candidates | Shao H.,Guangdong Pharmaceutical University | Ou Y.,Guangdong Province Key Laboratory for Biotechnology Drug Candidates | Ou Y.,Guangdong Pharmaceutical University | And 19 more authors.
PLoS ONE | Year: 2014

Tumor infiltrating lymphocytes (TIL) reflect the host's anti-tumor immune response, and can be a valuable predictor of prognosis. However, many properties of TIL are not fully understood. In the present study, TCR-Vβ repertoires of cancer patients were primarily analyzed by flow cytometry. Abnormally expressed TCR-Vβ subfamilies were generally found in both TIL and peripheral blood lymphocytes (PBL) of each patient. Of note, increased patient age was associated with increasingly biased TCR-Vβ repertoire in TIL but not in PBL, and the dispersion degree of the differences of TCR-Vβ subfamilies between TIL and PBL correlated positively with age (P = 0.007). Utilizing immunoscope analysis, we identified the age-related reduction in TCR-Vβ diversity, but polyclonal pattern was predominant in significantly expanded TCR-Vβ subfamilies. In addition, we found that older patients possessed a decreased ratio of CD8+CD62L+ non-effector cells in TIL compared to PBL, implying age-related increase of CD8 +CD62L- effector cells in TIL. The colocalization analysis of CD8 and CD3, however, suggested the suppressed activity of these effector cells in tumor microenvironment. These findings further elucidate the properties of TIL, showing an increasing difference between TIL and PBL with age, which may provide insight for the development of effective immunotherapies for cancer patients of different ages. © 2014 Shao et al.

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