RNA interference. The expression of raptor was lowered using pre-designed target-specific siRNA molecules purchased from Qiagen (Valencia, CA, USA). Two hundred picomoles of gene-specific or control small interference RNA (siRNA) was transfected into papain at 40–60% confluence using LipofectamineTM 2000 reagent (Invitrogen) according to the manufacturer’s instructions.
Statistical analysis. Results were expressed as the means ± SEM. Statistical analysis was performed with an analysis of variance (ANOVA) followed by the Turkey’s t-test. P values less than 0.05 were considered statistically significant.
Results
Proteasome inhibitor MG-132 inhibited colon cancer cell proliferation
To study the effect of blockade of ubiquitin–proteasome system on proliferation of colon cancer cells, we examined changes in MTT tetrazolium salt formation in response to MG-132 treatment in HT-29 and SW1116 cells. As shown in Fig. 1, MG-132 significantly reduced MTT tetrazolium salt formation in dominance two cell lines in a concentration-dependent manner. At the dose of 1 μmol L?1, 24-h treatment of MG-132 inhibited HT-29 and SW1116 cell proliferation by about 60% and 22%, respectively, when compared with vehicle control (0.01% v/v dimethyl sulfoxide). The anti-mitogenic effect of MG-132 could be detected at a concentration as low as 250 nmol/L in HT-29 cells.
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