Ku80 siRNA Silences Ku80 Expression and Enhances Radiosensitivity of A549 Lung Cancer Cells_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

REN Zhenyi , JIN Er , YE Jian , WANG Limin , QI Minghao

Department of Respiratory Medicine, The First Hospital of Hangzhou. Hangzhou, Zhejiang, 310006, ChinaCorresponding Author: REN Zhen-yi, E-mail: huxike123@ yahoo. com. cn;

Corresponding author：

Keywords：

A549 lung cancer cells; Ku80; siRNA interference; Radiosensitivity

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Objective To investigate the radiation-sensitizing effects of Ku80 silencing by siRNA interference for A549 lung cancer cells. Methods The sequences of Ku80 siRNA and negative siRNA were chemically synthesized and transfected into A549 lung cancer cells by lipofectamine. RT-PCR and Western bolt analysis were used to determine Ku80 gene expression. The transfected cells in culture dishes were irradiated with X ray at doses of 2 Gy, 4 Gy, 6 Gy, 8 Gy, 10 Gy, respectively. Once all treatments were completed, the cells were processed with the colony formation assay. Results RT-PCR detection showed that Ku80 mRNA levels in A549 lung cancer cells were reduced after transfected with Ku80 siRNA at 24 h, 48 h and 72 h time points. Western blot analysis showed that Ku80 protein content decreased at 48 h and 72 h time points compared with the control group ( P lt; 0. 05 ) . Cloning formation assay indicated that radiosensitivity of A549 lung cancer cells was enhanced after transfected with Ku80 siRNA. Conclusion Ku80 siRNA can effectively inhibit Ku80 gene expression of A549 lung cancer cells, and therefore enhance its radiosensitivity.

Citation： REN Zhenyi,JIN Er,YE Jian,WANG Limin,QI Minghao. Ku80 siRNA Silences Ku80 Expression and Enhances Radiosensitivity of A549 Lung Cancer Cells. Chinese Journal of Respiratory and Critical Care Medicine, 2010, 9(3): 291-294. doi: Copy

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.	金儿, 任振义, 卢忠, 等. X 线照射后A549 肺癌细胞Ku80 mRNA及其蛋白表达的变化. 中华放射医学与防护杂志, 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.	任振义, 白春学. 阿米福汀在肺癌放射治疗时对正常细胞和肺癌细胞的影响. 中国呼吸与危重监护杂志, 2005, 4: 181 -184.
6.	高阳, 李为民, 陈文彬. 抗hnRNP B1 单克隆抗体-顺铂交联物对人肺腺癌细胞A549 的体外靶向生长抑制作用. 中国呼吸与危重监护杂志, 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.	任振义, 金一尊. Ku 蛋白在肿瘤发生中的作用及其靶向抑制策略. 国际放射医学核医学杂志, 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.	管海涛, 薛兴欢, 王西京, 等. 靶向survivin 的siRNA 抑制乳腺癌MCF-7 细胞增殖并诱导凋亡. 中华肿瘤杂志, 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. 金儿, 任振义, 卢忠, 等. X 线照射后A549 肺癌细胞Ku80 mRNA及其蛋白表达的变化. 中华放射医学与防护杂志, 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. 任振义, 白春学. 阿米福汀在肺癌放射治疗时对正常细胞和肺癌细胞的影响. 中国呼吸与危重监护杂志, 2005, 4: 181 -184.
6. 高阳, 李为民, 陈文彬. 抗hnRNP B1 单克隆抗体-顺铂交联物对人肺腺癌细胞A549 的体外靶向生长抑制作用. 中国呼吸与危重监护杂志, 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. 任振义, 金一尊. Ku 蛋白在肿瘤发生中的作用及其靶向抑制策略. 国际放射医学核医学杂志, 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. 管海涛, 薛兴欢, 王西京, 等. 靶向survivin 的siRNA 抑制乳腺癌MCF-7 细胞增殖并诱导凋亡. 中华肿瘤杂志, 2006, 28: 326-330 .

Chinese Journal of Respiratory and Critical Care Medicine

Ku80 siRNA Silences Ku80 Expression and Enhances Radiosensitivity of A549 Lung Cancer Cells

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