| XIAO Wei,SUN Xin-chen,QIN Shu-kui,CHENG Hong-yan,ZHANG Wei,CAO Yuan-dong,LI Fan.Suppression of brain glioma cells expression of HMGA1 by lentivirus-mediated RNAi and its effect on radiosensitivity[J].Chinese Journal of Radiological Medicine and Protection,2010,30(2):129-132,142 |
| Suppression of brain glioma cells expression of HMGA1 by lentivirus-mediated RNAi and its effect on radiosensitivity |
| Received:October 29, 2009 |
| DOI： |
| KeyWords:RNA interference HMGA1 Human brain glioma Radiosensitivity |
| FundProject:国家自然科学基金(30970792);江苏省卫生厅科研基金(H200845);江苏省135工程医学重点学科 肿瘤放射治疗学基金(K0405) |
| Author Name | Affiliation | E-mail | | XIAO Wei | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | | | SUN Xin-chen | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | sunxch505@yahoo.com.cn | | QIN Shu-kui | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | | | CHENG Hong-yan | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | | | ZHANG Wei | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | | | CAO Yuan-dong | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | | | LI Fan | Department of Oncology, ZhongDa Hospital of Southeast University, Nanjing 210009,China | |
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| Abstract:: |
| Objective To investigate the radiosensitivity change of brain glioma cells of SHG-44 after HMGA1 inhibition by lentivirus-mediated RNA interference (RNAi).Methods Lentiviral vectors of HMGA1siRNA was constructed successfully and verified by PCR and DNA sequencing. After HMGA1siRNA was transfeced into the brain glioma cell line SHG-44, the expression of HMGA1 was determined by retrotranscriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. The effect of radiation exposure on cells was observed by clonogenic assay. Apoptosis and cell cycle were observed by flow cytometry.Results The results of RT-PCR and Western blot indicated that the expression of HMGA1 was down-regulated obviously in HMGA1 group. The expression of HMGA1 mRNA in HMGA1 group(0.11%±0.02%) was significantly less than that in untreated group(0.89%±0.02%, t=46.6, P＜0.01). The expression of HMGA1 protein in HMGA1 group(0.18%±0.02%)was also significantly decreased compared with that in untreated group(0.86%±0.03%, t=22.6, P＜0.01). Clonogenic assay showed that HMGA1 group was more sensitive than untreated group to 6 MV X-rays 12 d post-irradiation, with SER of 1.73. Flow cytometry demonstrated that the apoptosis rate of HMGA1 group (37.4%±3.1%) was higher than that of untreated group (6.1%±0.5%, t=12.9, P＜0.05). The rate of cell cycle delay in G0/ G1 was 72.6%±2.4% in HMGA1 group and 45.2%±1.6% in untreated group with significant statistical difference(t=16.2, P＜0.05).Conclusions RNAi vectors of HMGA1 could effectively suppress the expression of HMGA1 in SHG-44 cell line, so as to enhance the radio sensitivity of brain glioma cells. |
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