[Apoptosis and differentiation induced with TCLA in rat glioblastoma cells in vitro]
Ji-lei Hu1, Xiu-jie Wang, Pei-zhi Zhou
1Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To investigate the effect of conjugated trienoic fatty acids on the growth inhibition, apoptosis in rat glioblastoma cells and to elucidate its mechanism of activity.
Methods:
Rat glioblastoma cells were tested in vitro cytotoxicity, colony formation inhibition, Brdu incorporation after treatment with TCLA. Its effect of apoptosis induction was detected through Hoechst 33342 staining, cell cycle analysis. The expressions of ADPRTL1, CYP1A1 and PPAR-gamma genes were detected through RT-PCR.
Results:
After TCLA treatment, the proliferation of C6 cells were inhibited (40 micrommol/L, 72 h, viability 56.71% +/- 0.98%), this action acted as dose-time-dependent relations; colony formation decreased significantly (40 micrommol/L, 0) and BrdU labeling index of cancer cells decreased (63.1% +/- 1.0% vs 95.6 % +/- 1.4%); apoptotic cells increased; By FCM analysis, the apoptotic indices increased, the cells increased in G0/G1 phase, decreased in S phase, which have signigicant difference; RT-RCR showed that TCLA signigicantly increased the level of ADPRTL1, CYP1A1, and PPAR-gamma mRNA expression.
Conclusion:
The findings in this experimental study suggested that TCLA has potent cytotoxicity and induction apoptosis in human and rat glioblastoma cells, its mechanism of activity might be associated with the inhibition of DNA synthesis, cell cycle arrest, up-regulation the expression of apoptosis related genes ADPRTL, CYP1A1, PPAR-gamma.
