| GAO Ling,LI Feng-sheng,DONG Bo,et al.Effect of combination of STAT3 RNAi and 60Co γ-irradiation on U251 cell proliferation[J].Chinese Journal of Radiological Medicine and Protection,2008,28(1):17-20 |
| Effect of combination of STAT3 RNAi and 60Co γ-irradiation on U251 cell proliferation |
| Received:March 07, 2006 |
| DOI： |
| KeyWords:Small interfering RNA STAT3 U251 cell line |
| FundProject:国家自然科学基金资助项目(30770640) |
| Author Name | Affiliation | E-mail | | GAO Ling | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | LI Feng-sheng | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | DONG Bo | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | 张军权 | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | 饶亚岚 | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | 王治东 | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | 丛悦 | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | 毛秉智 | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | | | 陈肖华 | Beijing Institution of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China | chenxh@nic.bmi.ac.cn |
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| Abstract:: |
| Objective To construct signal transduction and activators of transcription 3 (STAT3) small interference RNA (siRNA) expression vector and to study its effect on STAT3 expression and U251 cell line proliferation.Methods STAT3 specific 19 bp oligonucleotides were designed and synthesized. These oligonucleotides were annealed to form the double strand DNA fragments and these fragments were cloned into Psilence2.1-U6-H1 vector. The recombinant of STAT3-siRNA expressing construction was confirmed by Hind Ⅲ and BamH Ⅰ double digestion and sequencing. The STAT3-siRNA was transfected into U251 cell. The inhibitory effect of STAT3-siRNA construction was tested by Western blot. Cellular proliferation activities were measured by tetrazolium bromide (MTT) colorimetry. Cloning efficiency and MTT were used to confirm the radiation dose.Results STAT3-siRNA expression vector was successfully constructed, and it could effectively down-regulate the protein levels of STAT3 in transfected U251 cell line;and the radiation dose was confirmed to 2 Gy. U251 cells transfected with STAT3-siRNA expression vector showed lower cellular proliferation compared with non-transfected U251 cells(P<0.05). Combination of 60Co γ-irradiation showed lower cellular proliferation compared with non-irradiated U251 cells(P<0.05). Conclusions The STAT3-siRNA expression vector can effectively inhibit STAT3 expression and gliosis cells proliferation. Combination with 2 Gy 60Co γ irradiation can enhance the inhibitory efficiency. |
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