题名：Adenovirus expressing shRNA to IGF-1R enhances the chemosensitivity of lung cancer cell lines by blocking IGF-1 pathway
作者：Yoon-Jin Lee, Arisa Imsumran, Mi-Young Park, Sung-Youn Kwon, Ho Il Yoon, Jae-Ho Lee, Chul-Gyu Yoo, Young Whan Kim, Sung Koo Han, Young-Soo Shim, et al.
来源：Lung Cancer[IF=3.172], Volume 55, Issue 3, March 2007, Pages 279-286
摘要： Yoon-Jin Leea, b, 1, Arisa Imsumranc, 1, Mi-Young Parka, b, Sung-Youn Kwona, b, Ho Il Yoona, b, Jae-Ho Leea, b, Chul-Gyu Yooa, Young Whan Kima, Sung Koo Hana, Young-Soo Shima, Wenhua Piaoc, Hiroyuki Yamamotoc, Yasushi Adachic, David P. Carboned and Choon-Taek Leea, b, ,
aDivision of Pulmonary and Critical Care Medicine, Department of Internal Medicine and Lung Institute of Medical Research Center, Seoul National University College of Medicine, Seoul 110-744, Republic of Korea
bDepartment of Medicine and Respiratory Center, Seoul National University Bundang Hospital, Seongnam 463-707, Republic of Korea
cFirst Department of Internal Medicine, Sapporo Medical University, Sapporo 060-8543, Japan
dVanderbilt-Ingram Cancer Center, Departments of Medicine and Cell Biology, Vanderbilt University School of Medicine, Nashville, TN 37232-6838, USA
Received 3 August 2006; revised 20 October 2006; accepted 23 October 2006. Available online 28 November 2006.
RNA interference is a phenomenon whereby small double-stranded RNA knocks down the expression of a sequence-specific gene. Double-stranded siRNA transfection, as currently used, is considered to have transient and low transfection efficiency. We constructed an adenoviral vector-based short hair-pin(sh)RNA system to overcome the limitations of the genetic blockade of IGF-1R, one of most important cancer therapy targets.
We constructed three different IGF-1R specific shRNAs (612, 801, and 3425) and generated three ad-shIGF-1Rs using BD Adeno-X? expression system. We assessed the effect of ad-shIGF-1R on signal transduction, induction of apoptosis, and in vitro tumorigenicity of lung cancer cell lines.
Western blot and FACS assays demonstrated that endogenous IGF-1R expression was efficiently suppressed after transduction of lung cancer cell lines with the three different ad-shIGF-1Rs. IGF-1R blockade by ad-shIGF-1R inhibited ligand induced phosphorylation of pAkt and pErk, and ad-shIGF-1R effectively blocked the in vitro tumorigenicity of lung cancer cell lines. Moreover, the transduction of a human lung cancer cell line with ad-IGF-1R(3425) enhanced chemosensitivity to anticancer drugs.
We conclude that the adenoviral vector-based approach to the RNA interference of IGF-1R induced effective IGF-1R silencing in lung cancer cell lines as manifested by effective blocking of the downstream pathway of IGF-1R and by an antitumor effect. We believe that this system can be usefully applied to other cancer targets. 32.