目的 探討用小RNA 干擾抑制Ku80 基因表達(dá)以提高A549 肺癌細(xì)胞的放射敏感性。方法 設(shè)計(jì)并化學(xué)合成Ku80 siRNA, 轉(zhuǎn)染體外培養(yǎng)的A549 肺癌細(xì)胞。利用RT-PCR 和Western blot分別從mRNA 和蛋白質(zhì)水平對轉(zhuǎn)染后的細(xì)胞Ku80 基因表達(dá)情況進(jìn)行測定, 同時(shí)上述轉(zhuǎn)染的細(xì)胞分別照射2、4、6、8 及10 Gy, 利用克隆形成實(shí)驗(yàn)測定放射敏感性的變化。結(jié)果 RT-PCR 檢測顯示在A549 肺癌細(xì)胞轉(zhuǎn)染Ku80 siRNA 后24、48 和72 h 三個(gè)時(shí)間點(diǎn)Ku80 mRNA 含量減少, Western blot 分析顯示在轉(zhuǎn)染48 和72 h 兩個(gè)時(shí)間點(diǎn)Ku80 蛋白含量減少, 與對照組比較有統(tǒng)計(jì)學(xué)差異( P lt;0. 05) 。克隆形成實(shí)驗(yàn)提示A549 肺癌細(xì)胞轉(zhuǎn)染Ku80 siRNA 后放射敏感性增強(qiáng)。結(jié)論 Ku80 siRNA 轉(zhuǎn)染A549 肺癌細(xì)胞后能夠有效抑制Ku80 基因表達(dá), 提高其放射敏感性。
引用本文： 任振義,金兒,葉健,王利民,祁明浩. 用Ku80 siRNA 抑制Ku80 基因表達(dá)以提高肺癌細(xì)胞放射敏感性的實(shí)驗(yàn)研究. 中國呼吸與危重監(jiān)護(hù)雜志, 2010, 9(3): 291-294. doi: 復(fù)制
| 1. | Sibanda BL, Chirgadze DY, Blundell TL. Crystal structure of DNAPKcs reveals a large open-ring cradle comprised of HEAT repeats.Nature, 2010, 463 : 118-121. |
| 2. | Holcomb VB, Rodier F, Choi Y, et al. Ku80 deletion suppresses spontaneous tumors and induces a p53-mediated DNA damage response. Cancer Res, 2008 , 68: 9497-9502. |
| 3. | 金兒, 任振義, 盧忠, 等. X 線照射后A549 肺癌細(xì)胞Ku80 mRNA及其蛋白表達(dá)的變化. 中華放射醫(yī)學(xué)與防護(hù)雜志, 2009, 29: 393 -395. |
| 4. | Yang QS, Gu JL, Du LQ, et al. ShRNA-mediated Ku80 gene silencing inhibits cell proliferation and sensitizes to gamma-radiation and mitomycin C-induced apoptosis in esophageal squamous cell carcinoma lines. J Radiat Res, 2008, 49: 399-407. |
| 5. | 任振義, 白春學(xué). 阿米福汀在肺癌放射治療時(shí)對正常細(xì)胞和肺癌細(xì)胞的影響. 中國呼吸與危重監(jiān)護(hù)雜志, 2005, 4: 181 -184. |
| 6. | 高陽, 李為民, 陳文彬. 抗hnRNP B1 單克隆抗體-順鉑交聯(lián)物對人肺腺癌細(xì)胞A549 的體外靶向生長抑制作用. 中國呼吸與危重監(jiān)護(hù)雜志, 2009 , 8: 451-455 . |
| 7. | Liu SK, Olive PL, Bristow RG. Biomarkers for DNA DSB inhibitors and radiotherapy clinical trials. Cancer Metastasis Rev, 2008, 27 :445-458. |
| 8. | Koike M, Koike A. Accumulation of Ku80 proteins at DNA doublestrand breaks in living cells. Exp Cell Res, 2008, 314: 1061-1070. |
| 9. | Steven AR, Dale AR. Loading of the nonhomologous end joining factor, Ku, on protein-occluded DNA ends. J Biol Chem, 2007, 282 :10605 -10613. |
| 10. | Diamanto M, Olivia N, Gerald BP, et al. Analysis of the DNA replication competence of the xrs-5 mutant cells defective in Ku86.J Cell Sci, 2003, 116: 111-124 . |
| 11. | 任振義, 金一尊. Ku 蛋白在腫瘤發(fā)生中的作用及其靶向抑制策略. 國際放射醫(yī)學(xué)核醫(yī)學(xué)雜志, 2008, 32: 305-307 . |
| 12. | Li GC, He F, Shao X, et al. Adenovirus-mediated heat-activated antisense Ku70 expression radiosensitizes tumor cells in vitro and in vivo. Cancer Res, 2003 , 63: 3268-3274. |
| 13. | Marangoni E, Foray N, O’Driscoii M, et al. A Ku80 fragment with dominant negative activity imparts a radiosensitive phenotype to CHO-K1 cells. Nuc Acids Res, 2000, 28: 4778-4782. |
| 14. | 管海濤, 薛興歡, 王西京, 等. 靶向survivin 的siRNA 抑制乳腺癌MCF-7 細(xì)胞增殖并誘導(dǎo)凋亡. 中華腫瘤雜志, 2006, 28: 326-330 . |
- 1. Sibanda BL, Chirgadze DY, Blundell TL. Crystal structure of DNAPKcs reveals a large open-ring cradle comprised of HEAT repeats.Nature, 2010, 463 : 118-121.
- 2. Holcomb VB, Rodier F, Choi Y, et al. Ku80 deletion suppresses spontaneous tumors and induces a p53-mediated DNA damage response. Cancer Res, 2008 , 68: 9497-9502.
- 3. 金兒, 任振義, 盧忠, 等. X 線照射后A549 肺癌細(xì)胞Ku80 mRNA及其蛋白表達(dá)的變化. 中華放射醫(yī)學(xué)與防護(hù)雜志, 2009, 29: 393 -395.
- 4. Yang QS, Gu JL, Du LQ, et al. ShRNA-mediated Ku80 gene silencing inhibits cell proliferation and sensitizes to gamma-radiation and mitomycin C-induced apoptosis in esophageal squamous cell carcinoma lines. J Radiat Res, 2008, 49: 399-407.
- 5. 任振義, 白春學(xué). 阿米福汀在肺癌放射治療時(shí)對正常細(xì)胞和肺癌細(xì)胞的影響. 中國呼吸與危重監(jiān)護(hù)雜志, 2005, 4: 181 -184.
- 6. 高陽, 李為民, 陳文彬. 抗hnRNP B1 單克隆抗體-順鉑交聯(lián)物對人肺腺癌細(xì)胞A549 的體外靶向生長抑制作用. 中國呼吸與危重監(jiān)護(hù)雜志, 2009 , 8: 451-455 .
- 7. Liu SK, Olive PL, Bristow RG. Biomarkers for DNA DSB inhibitors and radiotherapy clinical trials. Cancer Metastasis Rev, 2008, 27 :445-458.
- 8. Koike M, Koike A. Accumulation of Ku80 proteins at DNA doublestrand breaks in living cells. Exp Cell Res, 2008, 314: 1061-1070.
- 9. Steven AR, Dale AR. Loading of the nonhomologous end joining factor, Ku, on protein-occluded DNA ends. J Biol Chem, 2007, 282 :10605 -10613.
- 10. Diamanto M, Olivia N, Gerald BP, et al. Analysis of the DNA replication competence of the xrs-5 mutant cells defective in Ku86.J Cell Sci, 2003, 116: 111-124 .
- 11. 任振義, 金一尊. Ku 蛋白在腫瘤發(fā)生中的作用及其靶向抑制策略. 國際放射醫(yī)學(xué)核醫(yī)學(xué)雜志, 2008, 32: 305-307 .
- 12. Li GC, He F, Shao X, et al. Adenovirus-mediated heat-activated antisense Ku70 expression radiosensitizes tumor cells in vitro and in vivo. Cancer Res, 2003 , 63: 3268-3274.
- 13. Marangoni E, Foray N, O’Driscoii M, et al. A Ku80 fragment with dominant negative activity imparts a radiosensitive phenotype to CHO-K1 cells. Nuc Acids Res, 2000, 28: 4778-4782.
- 14. 管海濤, 薛興歡, 王西京, 等. 靶向survivin 的siRNA 抑制乳腺癌MCF-7 細(xì)胞增殖并誘導(dǎo)凋亡. 中華腫瘤雜志, 2006, 28: 326-330 .