The Role Of HSP70-2Gene In The Proliferation And Apoptosis Of Nasopharyngeal Carcinoma Cell

Objective:In this study, the HSP70-2gene expression of nasopharyngeal carcinoma cell6-10B could beup-regulated by Lentivirus technology. Then the HSP70-2gene expression of nasopharyngealcarcinoma cells6-10B or5-8F could be down-regulated by isoflavones drugs of ZSGS-4. Thereshould be explored the effects of HSP70-2gene on the proliferation of nasopharyngealcarcinoma cell.Methods:On the one hand, using of lentiviral technology the product was constructed into the targetgene lentiviral vector of pLV-HSP70-2-IRES/eGFP-Neo and the empty lentiviral vector ofpLV-CMV-eGFP-Neo. It could get the stably6-10B cell lines by the target gene lentivirus andthe empty lentivirus transducted respectively, i.e.6-10-HSP70-2/eGFP cells (the target genegroup) and6-10B-blank/eGFP cells (the blank control group). On the other hand, the HSP70-2gene expression of6-10B or5-8F cells was down-regulated by isoflavones drugs of ZSGS-4.The hsp70-2protein expression was analyzed by Western blotting, the proliferation ratios weredetected by CCK8and the cell cycle and the cell apoptosis were analyzed by flow cytometry.Results:In the first place, pEntry221-HSP70-2-IRES/eGFP entry vector was constructed. Then pLV-HSP70-2-IRES/eGFP-Neo lentiviral vector and pLV-CMV-eGFP-Neo empty vector wereconstructed successfully through LR recombination. In the second place, HSP70-2lentivirus waspacked by4lentiviral packaging plasmids system and the clonal cell clusters were screened byG418. hsp70-2protein expression of6-10B-HSP70-2/eGFP cells compared with that of6-10B-blank/eGFP cells or6-10B cells by Western blotting, then its level was respectivelyincreased by75.31±3.54%(P<0.01) or75.13±3.36%(P<0.01). It would be receivedsuccessfully6-10B-HSP70-2/eGFP and6-10B-blank/eGFP stably cells lines. The proliferationratio of6-10B-HSP70-2/eGFP cells compared with that of6-10B-blank/eGFP cells after24h,48h and72h of culture, its ratios were respectively increased by29.18±0.014%(P<0.01), 81.72±0.124%(P <0.001) and41.84±0.004%(P <0.001); or compared with that of6-10B cells,its ratios were respectively increased by25.69±0.031%(P<0.01),84.53±0.060%(P <0.001)and45.28±0.044%(P <0.001). PI and SPF of6-10B-HSP70-2/eGFP cells compared with thatof6-10B-blank/eGFP cells after24h of culture, its indexes were respectively increased by73.31±5.882%(P <0.001) and71.09±7.495%(P <0.01); or compared with that of6-10B cells, itsindexes were respectively increased by72.60±1.300%(P <0.001) and69.29±13.42%(P<0.01). Cell apoptosis ratios were not significantly changed among the three groups (P>0.05).For another, isoflavones drugs of ZSGS-4(0μg/mL,10μg/mL,12.5μg/mL,15μg/mL and17.5μg/mL) induced6-10B or5-8F with12h, hsp70-2protein expression was decreased(p<0.05), Inhibition ratios were significantly increased (p<0.05), DNA content of G0/G1phaseand S phase were significantly decreased (p<0.05), DNA content of SubG0/G1phase and G2/Mphase were significantly increased (p<0.05), Cell apoptosis ratios were significantly increased(p<0.05).Conclusion:The growth and proliferation of nasopharyngeal carcinoma cells6-10B would be promotedby up-regulated hsp70-2protein expression. The growth and proliferation of nasopharyngealcarcinoma cells6-10B or5-8F would be inhibited and apoptosis ratios were increased bydown-regulated hsp70-2protein expression. Hence HSP70-2gene might be effect on theproliferation of nasopharyngeal carcinoma cell.