Cell cycle distribution and induction of apoptosis of 95-D cells under AMDT treatment
To elucidate whether the decrease in cell viability induced by AMDT was attributable to apoptosis, nuclear staining with Hoechst 33258 and annexin V/PI staining were conducted.
By contrast, the condensation of nuclei and bleb-bing characteristic of apoptotic cells were evident in 95-D cells treated with different concentrations AMDT or positive control HCPT (Fig.
Table 2 Effects of AMDT on apoptosis and necrosis of 95-D human lung tumor cells (HCPT as positive control).
Effect of AMDT on mitochondrial transmembrane potential ([DELTA][psi]/m) and caspase-3 and -9 activities
To evaluate the influence of AMDT on the function of mitochondria, alterations in mitochondrial potential were analyzed by employing a fluorescent dye Rh123.
Following 24 h treatment of 95-D cells with various concentrations of AMDT, significant increases of both caspase-9 and -3 activities were detected and found to be in a dose-dependent manner compared with the control (Fig.
In this study, the new acyclic sesquiterpene AMDT from the hairy root cultures of A.
The growth inhibitory effect of AMDT on 95-D cells (Fig.
Further experiments revealed that apoptosis and cell cycle progression block were both responsible for the inhibition of the tumor cell proliferation by AMDT.
The results implied that AMDT might affect the G1/S cell-cycle checkpoint (Waldman et al.
These results demonstrated that AMDT efficiently induced apoptosis of 95-D cells.