| Abstract |
Objective: To establish a carboplatin (CBP)-resistant tongue squamous
carcinoma cell line, Tca8113/CBP cells, and to observe the reversal effect
of neferine in these cells.
Methods: Tca8113/CBP cell line was established by gradually increasing the
concentration of carboplatin and intermittent inducement to Tca8113 cells.
Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the reversal
effect of neferine on proliferation in Tca8113/CBP cells. mRNA expression
of drug-resistance-associated genes, lung related resistant protein
(LRP) and fibroblast muscle-type tropomyosin (FMT) were monitored by RT-PCR
in the cells before and after resistance establishment. Morphology of the
cells was observed under fluorescent microscopy, and the intracellular
changes of cytoskeleton between reversed Tca8113/CBP and non-reversed
Tca8113/CBP were compared.
Results: After 5 month's CBP treatment, Tca8113/CBP cells which can be
stably resistant to carboplatin (16 mug/ml) was successfully established.
The resistance index is 4.06. Neferine reversed the resistance of
Tca8113/CBP cells with a reversal fold of 2.12. Compared to the Tca8113
cells, the expression of LRP and FMT at mRNA level was increased obviously
in Tca8113/CBP cells. Whereas they were decreased after neferine treatment
(p<0.01), and the intracellular F-actin content was also decreased,
arranged in neat rows, more evenly distributed close to the level of
non-resistant Tca8113 cells.
Conclusions: Neferine exhibits the reversal effect on Tca8113/CBP cells,
which may be relate to decrease of LRP and FMT expression, increase of
intracellular content of carboplatin, change of the microfilament, reduce
of mechanical properties of adhesion, and interference of cell cycle in
Tca8113/CBP cells.
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