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Silencing RON gene expression via RNA interference inhibits human colon carcinoma HT-29 cell invasion and drug resistance
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Abstract:
Objective : To investigate the effect of receptor tyrosine kinase recepteur d’origine nantais (RON) gene silencing on the invasion and anticancer drug resistance of human colon carcinoma HT-29 cells. Methods: RNAi lentiviral vector targeting RON gene (Lv-RON-siRNA) was constructed. The efficiency of Lv-RON-siRNA on RON gene silence and RON protein level in HT-29 cells were detected by real-time PCR and Western blotting, respectively. The effects of Lv-RON-siRNA on invasion and drug resistance of HT-29 cells were observed by Transwell assay and ATP-TCA (ATP-tumor chemosensitivity assay). Results: The silencing effect of Lv-RON-siRNA on RON gene expression in HT-29 cells reached 70%. Compared with the control group, the invasion of HT-29 cells in Lv-RON-siRNA infection group was decreased（097±0.07 vs 1.29±0.08, P<0.05). The values of IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to 5-FU were (14.28±1.34) μg/ml and (8.93±1.2) μg/ml, respectively. The IC90 and IC50 of HT-29 cells infected with Lv-RON-siRNA to cisplatin (DDP) were (1.91±0.22) μg/ml and (0.64±0.07) μg/ml, respectively, and were significantly lower than those in the control group (P<0.01). Conclusion: Silencing RON gene expression can decrease the invasion ability of HT-29 cells and increase the sensitivity of HT-29 cells to 5-FU and DDP.
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Project Supported:
Project supported by the Natural Science Foundation of Ningbo City (No.2008A610082)
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