We aim to determine the regulation of apoptosis by paclitaxel-induced
and understand cancer dynamics to treatment targets for HeLa cells by identifying
decrease/increase genes expression on HeLa cells. In this study, the anti-tumor effects
of Paclitaxel (PAC) on HeLa cells have been studied in order to determine the
cellular and molecular mechanisms of these effects.
PAC has been applied to HeLa cells in 6 different doses (3, 7.5, 15, 30, 60, 120 nM)
for 48 hours and the IC50 dose MTT method, has been determined with apoptic index
(AI) DAPI. Morphological aspects have been demonstrated using light, phase
contrast and fluorescent microscopes, additionally activation of Caspase 3,7 and 10
have been shown using florescent spectroscopy. RT-PCR and qRT-PCR have been
used to evaluate pro/anti-apoptotic gene expression.
According to the parameters being evaluated; PAC has reduced cell multiplication based on dosage and
time (p<0.01). 15 nM has been determined as the IC50 value. AI value has been determined as 42%. In
the molecular level analyses in addition to the increase in Caspase3,7,10 activation, RT-PCR results
show that bax, bak, bcl-x, bik, mcl-1 genes are expressed in the control group as well as the experimental
15 nM group; whereas bak, bcl-x ve bik genes have a decrease in expression compared to the control
group. qRT-PCR results show that Apaf1, Bad, Bax, Bcl2L11, Caspase1, Caspase10, Caspase4,
Caspase7, Dffa, Fas, Htra2, Lrdd, NFKB1, NFKB2, PMAIP1, RELA, RELB, TNFRSF10A,
TNFRSF10C, TNFRSF10D, TNFRSF1A, TNFRSF21, TNFRSF25 gene expressions have increased
significantly. On the other hand, BAG1, BBC3, Bcl2L1, Bcl2L10, Bid, Caspase2, Caspase6, Caspase8,
Caspase9, FADD, FAM96A, FasLG, HRK, SOCS3, TNF, TNFSF10, TRAF5, TRAF6 mRNA levels
are significantly decreased.