| WANG Pei-guo,LIU Zhi-yan,WEI feng.Experimental study of antisense epidermal growth factor receptor enhancing the radiosensitivity of human lung cancer cell line spc-a-1[J].Chinese Journal of Radiological Medicine and Protection,2008,28(4):361-364 |
| Experimental study of antisense epidermal growth factor receptor enhancing the radiosensitivity of human lung cancer cell line spc-a-1 |
| Received:December 26, 2007 |
| DOI： |
| KeyWords:Lung adenocarcinoma Antisense EGFR Radiosensitivity |
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| Abstract:: |
| Objective To explore whether antisense-EGFR could enhance the radiosensitivity of human lung cancer spc-a-1 cell line. Methods The spc-a-1 cells were transfected with antisense-EGFR-pcDNA3 by lipofectamine 2000(pcDNA3 antiEGFR group). Two other groups were used for comparison: control group (spc-a-1 cell without transfection) and pcDNA3 group (spc-a-1 cell transfected with pcDNA3 which did not contain antisense EGFR). Cell clones that stable expressing antisense-EGFR was selected with G418 and the suppression of the expression of EGFR mRNA and protein were detected by RT-PCR and Western blot. The influence of antisense-EGFR on cell cycle was testified by flow cytometry assay. The cell apoptosis was analyzed by flow cytometry after 8 Gy irradiation. Further, cells of each group were irradiated with X-rays at the dose of 0, 2, 4, 6 and 8 Gy. Dose-survival curve of each group was established by colony-forming assay. Results The expression of EGFR mRNA and protein were significantly inhibited after antisense-EGFR- pcDNA3 transfection. The cells arrested at the G2/M phase in the pcDNA3 antiEGFR group, control group and pcDNA3 group were (29.53±1.91)％, (13.7±1.30)% and (12.40±1.34)％,respectively. The apoptosis index of spc-a-1 cells in the antisense-EGFR combined with irradiation group was obviously higher than that of the comparable groups [(39.24±1.57)%, (13.79±0.63)% and (15.02±0.85%)]. The values of D0, Dq, SF2 of pcDNA3 antiEGFR group declined obviously compared with the control group(2.11, 2.49, 0.84 vs 1.19, 0.15, 0.32).Conclusions Antisense-EGFR could induce the G2/M cell cycle arrest, promote cell apoptosis and inhibit the ability of sublethal cell damage repair induced by irradiation, so that it could significantly improve the radiosensitivity of spc-a-1 cell in vitro. |
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