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miRNA-200c enhances sensitivity of gastric cancer SGC7901/DDP cells to cisplatin through inhibiting Akt pathway
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Abstract:
Objective:To explore the effect of miRNA-200c on chemosensitivity of drug-resistant human gastric cancer SGC7901/DDP cells to cisplatin (DDP) and its mechanism. Methods: The expressions of E-cadherin, PTEN, p-Akt and total Akt proteins in SGC7901/DDP cells and its parental SGC7901 cells were detected by Western blotting. miRNA-200c precursor (Pre-200c) was transiently transfected into SGC7901/DDP cells to increase the expression of miRNA-200c. The drug sensitivity of cells to cisplatin and the expression level of p-Akt were detected by MTT assay and Western blotting, respectively. Finally, SGC7901/DDP cells were treated with LY94002 to inactivate the phosphorylation of Akt, and the sensitivity of SGC7901/DDP cells to DDP was detected. Results: Compared with SGC7901 cells, the expression of p-Akt protein in SGC7901/DDP cells was significantly increased (\[1.02±0.09\] vs \[0.17±0.02\], P<0.05), while the expressions of E-cadherin and PTEN protein were significantly decreased (\[0.10±0.03\] vs \[0.47±0.06\]; \[0.18±006\] vs \[0.87±0.06\] respectively, P<0.05). The IC50 of cisplatin in the Pre-200c tansfected group was significantly lower than that in the negative control group (\[7.52±0.19\] mg/L vs \[12.18±0.29\] mg/L, P<0.05). Furthermore, the expression of p-Akt protein in the Pre-200c transfected group was significantly lower than that in the control group (\[0.22±0.04\] vs \[0.69±0.09\], P<0.05); the level of p-Akt protein was significantly inhibited (\[0.18±0.06\] vs \[0.66±0.10\], P<0.05) in SGC7901/DDP cells treated with LY94002. Moreover, the IC50 of DDP in the LY94002 treated group was significantly lower than that in the control group (\[6.80±0.28\] mg/L vs \[11.94±1.73\] mg/L, P<0.05). Conclusion: Drug resistance phenotype of SGC7901/DDP cells may be associated with the loss of E-cadherin and PTEN proteins and abnormally activation of Akt signaling pathway, and the chemotherapeutic sensitization of miRNA-200c may be through the inactivation of Akt signal pathway.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81172333)
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