Abstract
The type-1 insulin-like growth factor receptor (IGF-1R) is over-expressed by endometrial carcinoma, level of IGF-1R has been correlated with tumor progression, and high IGF-1R expression has been found to be an important prognostic factor. In the study, we used lentivirus-mediated shRNA targeting IGF-1R to silence its expression, then assessed the effect of down-regulation of this receptor on cell growth and chemosensitivity to cisplatin. Lentivirus-mediate shRNA was designed and transfected to the endometrial carcinoma HEC-1B cell. The IGF-1R mRNA and related protein expression, cell proliferation ability, cell apoptosis, and cell cycle change were detected. Cell proliferation inhibition rates, cell apoptosis, and level of cleaved caspase-9 were measured in various concentrations of cisplatin. The mRNA and protein level of IGF-1R, and the phosphorylated protein p-Akt, p-Erk were all suppressed after transfection. Cell proliferation was inhibited in successive five days after transfection, the highest inhibition rate was 43.28 ± 3.55% on day 5. After transfection, 24.96 ± 1.05% cells were in G2/M phase, and cell apoptotic rate increased from 10.66 ± 0.08 to 19.92 ± 1.34%. In various concentrations of cisplatin, transfected cells proliferation was significantly inhibited which made the IC50 value drop from 21.85 uM to 10.58 uM. Incubation with different concentrations of cisplatin for 48 h, cells apoptotic rate increased to 41.92 ± 2.5, 31.13 ± 2.76, 22.21 ± 4.63%, respectively, which was accompanied with increased cleaved caspase-9 expression. Lentivirus-mediated shRNA targeting IGF-1R has the potential to develop as a clinical treatment method in advanced and chemoresistant endometrial carcinoma.
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References
Kurman RJ, Zaino RJ, Norris HJ (1994) Endometrial carcinoma. In: Kurmann RJ (ed) Blaustein’s pathology of the female genital tract, 4th edn. Springer Verlag, New York, pp 447–448
Barakat RR, Grisby PW, Sabbatini P (2000) Corpus:epithelial tumors. In: Hoskins WJ, Perez CA, Young RC (eds) Principles and practice of gynecologic oncology, 2nd edn. Lippincott Williams and Wilkins, Philadelphia, pp 919–959
Carey MS, Gawlik C, Fung-Kee-Fung M, Chambers A, Oliver T (2006) Systematic review of systemic therapy for advanced or recurrent endometrial cancer. Gynecol Oncol 101:158–167
Werner H, Bruchim I (2009) The insulin-like growth factor-I receptor as an oncogene. Arch Physiol Biochem 115:58–71
Yavari K, Taghikhani M, Maragheh MG, Mesbah-Namin SA, Babaei MH (2009) Knockdown of IGF-IR by RNAi inhibits SW480 colon cancer cells growth in vitro. Arch Med Res 40:235–240
Sachdev D, Hartell JS, Lee AV, Zhang X, Yee D (2004) A dominant negative type I insulin-like growth factor receptor inhibits metastasis of human cancer cells. J Biol Chem 279:5017–5024
Wolf S, Lorenz J, Mossner J, Wiedmann M (2010) Treatment of biliary tract cancer with NVP- AEW541: mechanisms of action and resistance. World J Gastroenterol 16:156–166
Furukawa J, Wraight CJ, Freier SM, Peralta E, Atley LM, Monia BP, Gleav ME, Cox ME (2010) Antisense oligonucleotide targeting of insulin-like growth factor-1 receptor (IGF-1R) in prostate cancer. Prostate 70:206–218
Yavari K, Taghikhani M, Maragheh MG, Mesbah-Namin SA, Babaei MH, Arfaee AJ, Madani H, Mi-rzaei HR (2010) SiRNA-mediated IGF-1R inhibition sensitizes human colon cancer SW480 cells to radiation. Acta Oncol 49:70–75
Wang YH, Xiong J, Wang SF, Yu Y, Wang B, Chen YX, Shi HF, Qiu Y (2010) Lentivirus mediated shRNA targeting insulin-like growth factor-1 receptor (IGF-1R) enhances chemosensitivity of osteosarcoma cells in vitro and in vivo. Mol Cell Biochem 341:225–233
Larsson O, Girnita A, Girnita L (2005) Role of insulin-like growth factor 1 receptor signalling in cancer. Br J Cancer 92:2097–2101
Ryan PD, Goss PE (2008) The emerging role of the insulin-like growth factor pathway as a therapeutic target in cancer. Oncologist 13:16–24
Pavelić J, Radaković B, Pavelić K (2007) The insulin-like growth factor-2 and its receptors (IGF-1R and IGF-2R/mannose6-phosphate) in endometrial adenocarcinoma. Gynecol Oncol 105:727–735
Kurihara S, Hakuno F, Takahashi S (2000) Insulin-like growth factor-I-dependent signal transduction pathways leading to the induction of cell growth and differentiation of human neuroblastoma cell line SH-SY5Y: the roles of MAP kinase pathway and PI3-kinase pathway. Endocr J 47:739–751
Pollak M (2008) Targeting insulin and insulin-like growth factor signalling in oncology. Curr Opin Pharmacol 8:384–392
Pollak M (2008) Insulin, insulin-like growth factors and neoplasia. Best Pract Res Clin Endocrinol Metab 22:625–638
Pfeffer S (2010) RNA silencing as a natural antiviral defense system in mammals: where are we now? Mol Ther 18:871–872
Downward J (2004) RNA interference. BMJ 328:1245–1248
Sumimoto H, Kawakami Y (2010) The RNA silencing technology applied by lentiviral vectors in oncology. Method Mol Biol 614:187–199
Shu SR, Li XM, Yang YB, Zhang Y, Li T, Liang CY, Wang J (2010) Inhibitory effect of siRNA targeting IGF-1R on endometrial carcinoma. Int Immunopharmacol 11:244–249
Dong AQ, Kong MJ, Ma ZY (2007) Down-regulation of IGF-IR using small, interfering, hairpin RNA (siRNA) inhibits growth of human lung cancer cell line A549 in vitro and in nude mice. Cell Biol Int 31:500–507
LeRoith D, Helman L (2004) The new kid on the block (ade) of the IGF-1 receptor. Cancer Cell 5:201–202
Niu J, Xu Z, Li XN, Han Z (2007) siRNA-mediated type 1 insulin-like growth factor receptor silencing induces chemosensitization of a human liver cancer cell line with mutant P53. Cell Biol Int 31:156–164
Maloney EK, McLaughlin JL, Dagdigian NE, Garrett LM, Connors KM, Zhou XM, Blattler WA, Chittenden T, Singh R (2003) An anti-insulin-like growth factor I receptor antibody that is a potent inhibitor of cancer cell proliferation. Cancer Res 63:5073–5083
Hellawell GO, Ferguson DJ, Brewster SF, Macaulay VM (2003) Chemosensitization of human prostate cancer using antisense agents targeting the type 1 insulin-like growth factor receptor. Br J Urol 91:271–277
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This work was supported by the Science and Technology Plan Project of Guang Dong Province (2007B 030502014, 00429391120223052) and the National Nature Science Foundation of China (30772332).
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Shu, S., Yang, Y., Li, X. et al. Down-regulation of IGF-1R expression inhibits growth and enhances chemosensitivity of endometrial carcinoma in vitro. Mol Cell Biochem 353, 225–233 (2011). https://doi.org/10.1007/s11010-011-0790-9
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DOI: https://doi.org/10.1007/s11010-011-0790-9