Effects Of Nuclear M-csf On The Sensitivity Of Anti-tumor Chemotherapeutic Drug In Human Cervical Carcinoma Hela Cells

OBJECTIVETo investigate effects of nuclear M-CSF on the sensitivity of anti-tumor chemotherapeutic drug in human cervical carcinoma HeLa cells.METHODSHuman cervical carcinoma HeLa cell, HeLa-M cell (pCMV/cyto/myc-M-CSF transfected-HeLa cell that highly expresses nuclear M-CSF, referred as to HeLa-M cell) and HeLa-C cell (vector pCMV/cyto/myc transfected HeLa cell, referred as to HeLa-C cell) were cultured for 48 h or 2 h. The cell viability was determined using MTT and Trypan blue assay. The apopotosis percentage was examined by flow cytometry using Annexin V/PI fluorescence staining. The concentration of intracellular cisplatin or paclitaxel was detected with high performance liquid chromatogram(HPLC).The expression of Bcl-2 protein was analyzed by Western blot.RESULTSThe MTT and Trypanblue assay showed that survival rate of HeLa-M cell markedly higher than that of both HeLa-C cell and HeLa cell (p<0.01) when these cell were treated with a concentration of 2.0μg/mL or 20.0μg/mL of either cisplatin or paclitaxel. There was no defference for survival rate in the three kind of cells (p>0.05) when cells were incubated with a concentration of 0.2μg/ mL of these two drugs. Annexin V/PI fluorescence staining and FCM indicated that apopotosis percentage of HeLa-M cells was remarkably lower than that of both HeLa-C cell and HeLa cell treated with 2.0μg/mL or 20.0μg/mL(p<0.01), but not 0.2μg/ mL(p>0.05) of either cisplatin or paclitaxel for 2 h. HPLC assay showed that the concentration of intracellular cisplatin or paclitaxel of HeLa-M cell was significantly lower than that of both HeLa-C cell and HeLa cell(p<0.01) when cell treated with 2.0μg/mL or 20.0μg/mL(p<0.01), but not 0.2μg/ mL(p>0.05) of cisplatin or paclitaxel. There was no defference for intracellular cisplatin or paclitaxel in the three kind of cells (p>0.05) . Western blot indicated that Bcl-2 expression in HeLa-M cell is more heightened than that in HeLa cell or HeLa-C cell (p<0.01).CONCLUSION1. Nuclear M-CSF reinforces the drug resistance to both cisplatin and paclitaxel and reduces the concentration of these two drugs within human cervical carcinoma HeLa cell.2. Nuclear M-CSF up-regulates Bcl-2 expression and enhances the anti-apoptosis ability of human cervical carcinoma HeLa cell.