| Summary: | Objective To explore the effect of sodium hydroxide(NaOH) and rapamycin(Rapa) alone or their combination in treating Tsc2-null(Tsc2-/-) mouse embryonic fibroblasts. Methods The wild type and Tsc2-/- cells were treated with NaOH and/or Rapa and divided into 4 groups: control, NaOH group, Rapa group, and combination (Combo) group. Cell proliferation was measured by MTT assay and colony formation. Flow cytometry was used to detect the potential influence on cell cycle. The protein expression of S6, p-S6, p-4EBP1 was detected by Western blot. Results Compared to control group, Tsc2-/- cells in NaOH and Rapa groups both showed a reduced proliferation ability and colony counting (P＜0.05); additionally, only in Rapa group, the percentage of Tsc2-/- cells in G1 phase increased and the protein level of p-S6 and p-4EBP1 all decreased (P＜0.05). Compared with NaOH and Rapa groups, Tsc2-/- cells in Combo group had a further decreased in proliferation ability and colony number and an increased G1 phase percentage (P＜0.05). But the level of p-S6 and p-4EBP1 of Tsc2-/- cells in Combo group was significantly lower than those in NaOH group (P＜0.05). Conclusions NaoH(15 μmol/L) enhances the suppressing poliferation efficacy of Rapa in Tsc2-/- cells.
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