Abstract
BackgroundRNA interference (RNAi) is a powerful approach in functional genomics to selectively silence messenger mRNA (mRNA) expression and can be employed to rapidly develop potential novel drugs against a complex disease like cancer. However, naked siRNA being anionic is unable to cross the anionic cell membrane through passive diffusion and therefore, delivery of siRNA remains a major hurdle to overcome before the potential of siRNA technology can fully be exploited in cancer. pH-sensitive carbonate apatite has recently been developed as an efficient tool to deliver siRNA into the mammalian cells by virtue of its high affinity interaction with the siRNA and the desirable size distribution of the resulting siRNA-apatite complex for effective cellular endocytosis. Moreover, internalized siRNA was found to escape from the endosomes in a time-dependent manner and efficiently silence gene expression.ResultsHere we show that carbonate apatite-mediated delivery of siRNA against PLC-gamma-2 (PLCG2) and calmodulin 1 (CALM1) genes has led to the sensitization of a human cervical cancer cell line to doxorubicin- and paclitaxel depending on the dosage of the individual drug whereas no such enhancement in cell death was observed with cisplatin irrespective of the dosage following intracellular delivery of the siRNAs.ConclusionThus, PLCG2 and CALM1 genes are two potential targets for gene knockdown in doxorubicin and paclitaxel-based chemotherapy of cervical cancer.
Highlights
RNA interference (RNAi) is a powerful approach in functional genomics to selectively silence messenger mRNA expression and can be employed to rapidly develop potential novel drugs against a complex disease like cancer
We show that carbonate apatite-mediated delivery of Small interfering RNA (siRNA) against phospholipase C (PLC)-gamma-2 (PLCG2) and calmodulin 1 (CALM1) genes sensitized a human cervical cancer cell line (HeLa cell) to doxorubicin- and paclitaxelinduced cell death depending on the doses of the drugs while no such synergistic effect was observed with cisplatin, another commonly used chemotherapy drugs
Roles of PLCG2 and CALM1 in the proliferation/survival of cervical cancer cells In order to investigate the potential roles of PLCG2 and CALM1 in the proliferation or survival of HeLa cells that express both of the proteins [20,21,22,23], specific validated siRNA (10 mM) against PLCG2 or CALM1 mRNA was added together with Ca2+ (3 mM) to the bicarbonate-buffered Dulbecco's Modified Eagle’s Medium (DMEM) prior to the incubation at 37 °C for 30 min to form carbonate apatite/siRNA complexes
Summary
We show that carbonate apatite-mediated delivery of siRNA against PLC-gamma-2 (PLCG2) and calmodulin 1 (CALM1) genes has led to the sensitization of a human cervical cancer cell line to doxorubicin- and paclitaxel depending on the dosage of the individual drug whereas no such enhancement in cell death was observed with cisplatin irrespective of the dosage following intracellular delivery of the siRNAs
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