Downregulation of Tetraspanin 8 and Carbonic Anhydrase 9 gene expression sensitized pancreatic cancer cells to cisplatin.

Abstract

In this context, we used the combined siRNA and chemotherapeutic drug treatment strategy for minimizing the side effects of chemotherapy agents in pancreatic cancers. We used the gene silencing approach with siRNA to target CA9 and TSPAN8 genes, which are overexpressed in pancreatic cancer. Then expression of the targeted CA9 and TSPAN8 genes were determined by western blot analysis. Effect gene silencing on the proliferation of Panc-1 and MiaPaca-2 cells were detected by MTT assay after transfection with Control siRNA, CA9 siRNA, and TSPAN8 siRNA. MTT cell proliferation assay was also performed to determine the effect the combined siRNA and chemotherapeutic drug treatment as to evaluate cisplatin and epirubicin sensitivity. The results demonstrated that targeted CA9 siRNA and TSPAN8 siRNA have increased the sensitivity of pancreatic cells to cisplatin and epirubicin that can conclude; CA9 and TSPAN8 silencing may be a suitable candidate for therapeutic applications.

Keywords

• CA9,TSPAN8,siRNA,pancreatic cancer

References

1. 1. Del Chiaro, M., et al.,2014. Early detection and prevention of pancreatic cancer: is it really possible today? World J Gastroenterol. 20(34): p. 12118-31.
2. 2. Shi, S., et al.,2012. Combinational therapy: new hope for pancreatic cancer? Cancer Lett. 317(2): p. 127-35.
3. 3. El Maalouf, G., et al.,2009. Markers involved in resistance to cytotoxics and targeted therapeutics in pancreatic cancer. Cancer Treat Rev. 35(2): p. 167-74.
4. 4. Saraswathy, M. and S. Gong,2014. Recent developments in the co-delivery of siRNA and small molecule anticancer drugs for cancer treatment. Materials Today. 17(6): p. 298-306.
5. 5. Kanetaka, K., et al.,2001. Overexpression of tetraspanin CO-029 in hepatocellular carcinoma. J Hepatol. 35(5): p. 637-42.
6. 6. Zoller, M.,2009. Tetraspanins: push and pull in suppressing and promoting metastasis. Nat Rev Cancer. 9(1): p. 40-55.
7. 7. Kim, T.K., et al.,2015. Generation of a human antibody that inhibits TSPAN8-mediated invasion of metastatic colorectal cancer cells. Biochem Biophys Res Commun. 468(4): p. 774-80.
8. 8. Berthier-Vergnes, O., et al.,2011. Gene expression profiles of human melanoma cells with different invasive potential reveal TSPAN8 as a novel mediator of invasion. Br J Cancer. 104(1): p. 155-65.

9. 9. Wei, L., Y. Li, and Z. Suo,2015. TSPAN8 promotes gastric cancer growth and metastasis via ERK MAPK pathway. Int J Clin Exp Med. 8(6): p. 8599-607.
10. 10. Fang, T., et al.,2016. Tetraspanin-8 promotes hepatocellular carcinoma metastasis by increasing ADAM12m expression. Oncotarget. 7(26): p. 40630-40643.
11. 11. Akiel, M.A., et al.,2016. Tetraspanin 8 mediates AEG-1-induced invasion and metastasis in hepatocellular carcinoma cells. FEBS Lett. 590(16): p. 2700-8.
12. 12. Park, C.S., et al.,2016. Therapeutic targeting of tetraspanin8 in epithelial ovarian cancer invasion and metastasis. Oncogene. 35(34): p. 4540-8.
13. 13. Huang, W.J., et al.,2015. Expression of hypoxic marker carbonic anhydrase IX predicts poor prognosis in resectable hepatocellular carcinoma. PLoS One. 10(3): p. e0119181.
14. 14. Kazokaite, J., et al.,2017. An update on anticancer drug development and delivery targeting carbonic anhydrase IX. PeerJ. 5: p. e4068.
15. 15. Yin, T., et al.,2015. Co-delivery of hydrophobic paclitaxel and hydrophilic AURKA specific siRNA by redox-sensitive micelles for effective treatment of breast cancer. Biomaterials. 61: p. 10-25.
16. 16. Lee, S.Y., et al.,2016. A theranostic micelleplex co-delivering SN-38 and VEGF siRNA for colorectal cancer therapy. Biomaterials. 86: p. 92-105.
17. 17. Svastova, E., et al.,2012. Carbonic anhydrase IX interacts with bicarbonate transporters in lamellipodia and increases cell migration via its catalytic domain. J Biol Chem. 287(5): p. 3392-402.
18. 18. Csaderova, L., et al.,2013. The effect of carbonic anhydrase IX on focal contacts during cell spreading and migration. Front Physiol. 4: p. 271.
19. 19. Gomes, I.M., et al.,2018. Knockdown of STEAP1 inhibits cell growth and induces apoptosis in LNCaP prostate cancer cells counteracting the effect of androgens. Med Oncol. 35(3): p. 40.
20. 20. Whitehurst, A.W., et al.,2007. Synthetic lethal screen identification of chemosensitizer loci in cancer cells. Nature. 446(7137): p. 815-9.
21. 21. Mezencev, R., et al. Acquired resistance of pancreatic cancer cells to cisplatin is multifactorial with cell context-dependent involvement of resistance genes. Cancer Gene Ther. 23(12): p.446–453.
22. 22. Plosker, G.L. and D. Faulds. Epirubicin. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic use in cancer chemotherapy. Drugs. 45(5):788-856.
23. 23. Beh, C.W., et al.,2009. Efficient delivery of Bcl-2-targeted siRNA using cationic polymer nanoparticles: downregulating mRNA expression level and sensitizing cancer cells to anticancer drug. Biomacromolecules. 10(1): p. 41-8.
24. 24. Chen, A.M., et al.,2009. Co-delivery of doxorubicin and Bcl-2 siRNA by mesoporous silica nanoparticles enhances the efficacy of chemotherapy in multidrug-resistant cancer cells. Small. 5(23): p. 2673-7.
25. 25. Yang, Z.Z., et al.,2014. Tumor-targeting dual peptides-modified cationic liposomes for delivery of siRNA and docetaxel to gliomas. Biomaterials. 35(19): p. 5226-39.