Ginkgetin induces apoptosis in 786-O cell line via suppression of JAK2-STAT3 pathway

Document Type: Original Article


Laboratory of kidney Carcinoma, Ningbo Urology and Nephrology Hospital, Urology and Nephrology Institute of Ningbo University, Ningbo 315000, Zhejiang Province, P.R. China


Objective(s): Renal cell carcinoma (RCC) is insensitive to conventional chemotherapy. Ginkgetin effectively treats several carcinoma cells. However, little is known about effects of Ginkgetin on RCC. In the present study, using 786-O cells, we evaluate whether Ginkgetin exerts anticancer effects against RCC.
Materials and Methods: 786-O cells suspended in the medium containing Ginkgetin were cultured for 24 hr to 72 hr, and then MTT assay was used to study cytotoxic effect of Ginkgetin. Apoptosis in 786-O was measured by an FITC Annexin apoptosis detection kit. Protein expression was detected by Western blotting. 786-O cells with active Janus kinase 2 (JAK2)-Signal transducer and activator of transcription 3 (STAT3) were prepared by stimulant of interleukin-6 (IL-6), whereas 786-O cells with deactivated STAT3 were produced by small interfering RNA (siRNA) STAT3.
Results: Ginkgetin suppressed the growth of 786-O in dose and time-dependent manners with IC50 values of 7.23 μM. Ginkgetin induced apoptosis of 786-O cells and increased the levels of caspase-8, caspase-9, and caspase-3. Additionally, Ginkgetin treated 786-O cells showed decreased levels of JAK2 and phosphorylated-STAT3 whether or not IL-6 was pretreated. Interestingly, pretreatment of siRNA STAT3 exerted inhibitory effects on the growth of 786-O cells, and the observation could be further reinforced after the Ginkgetin treatment.
Conclusion: Our results indicate Ginkgetin possesses obvious inhibitory effects on the proliferation of 786-O, and this effect is probably due to its inhibition of JAK2/STAT3 pathway. Our findings imply Ginkgetin is a potential therapeutic medicine for RCC.


1. Ljungberg B, Campbell SC, Choi HY, Jacqmin D, Lee JE, Weikert S, et al. The epidemiology of renal cell carcinoma. Eur Urol 2011; 60:615-621.

2. Ridge CA, Pua BB, Madoff DC. Epidemiology and staging of renal cell carcinoma. Semin Intervent Radiol 2014; 31:3-8.

3. Fang Z, Tang Y, Fang J, Zhou Z, Xing Z, Guo Z, et al. Simvastatin inhibits renal cancer cell growth and metastasis via AKT/mTOR, ERK and JAK2/ STAT3 pathway. PLoS One 2013; 8:e62823.

4. Berger A, Kamoi K, Gill IS, Aron M. Cryoablation for renal tumors: current status. Curr Opin Urol 2009; 19:138-142.

5. Mukthavaram R, Ouyang X, Saklecha R, Jiang P, Nomura N, Pingle SC, et al. Effect of the JAK2/STAT3 inhibitor SAR317461 on human glioblastoma tumorspheres. J Transl Med 2015; 13:269.

6. Wu L, Guo L, Liang Y, Liu X, Jiang L, Wang L. Curcumin suppresses stem-like traits of lung cancer cells via inhibiting the JAK2/STAT3 signaling pathway. Oncol Rep 2015; 34:3311-3317.

7. Liu YF, Lu YM, Qu GQ, Liu Y, Chen WX, Liao XH, et al. Ponicidin induces apoptosis via JAK2 and STAT3 signaling pathways in gastric carcinoma. Int J Mol Sci 2015; 16:1576-1589.

8. Jorvig JE, Chakraborty A. Zerumbone inhibits growth of hormone refractory prostate cancer cells by inhibiting JAK2/STAT3 pathway and increases paclitaxel sensitivity. Anticancer Drugs 2015; 26:160-166.

9. Lim H, Son KH, Chang HW, Kang SS, Kim HP. Effects of anti-inflammatory biflavonoid, ginkgetin, on chronic skin inflammation. Biol Pharm Bull 2006; 29:1046-1049.

10. Miki K, Nagai T, Suzuki K, Tsujimura R, Koyama K, Kinoshita K, et al. Anti-influenza virus activity of biflavonoids. Bioorg Med Chem Lett 2007; 17:772-775.

11. Wang YQ, Wang MY, Fu XR, Peng-Yu, Gao GF, Fan YM, et al. Neuroprotective effects of ginkgetin against neuroinjury in Parkinson's disease model induced by MPTP via chelating iron. Free Radic Res 2015; 49:1069-1080.

12. Su Y, Sun CM, Chuang HH, Chang PT. Studies on the cytotoxic mechanisms of ginkgetin in a human ovarian adenocarcinoma cell line.Naunyn Schmiedebergs Arch Pharmacol 2000; 362:82- 90.

13. You OH, Kim SH, Kim B, Sohn EJ, Lee HJ, Shim BS, et al. Ginkgetin induces apoptosis via activation of caspase and inhibition of survival genes in PC-3 prostate cancer cells. Bioorg Med Chem Lett 2013; 23:2692-2695.

14. Ye ZN, Yu MY, Kong LM, Wang WH, Yang YF, Liu JQ, et al. Biflavone Ginkgetin, a Novel Wnt Inhibitor, Suppresses the Growth of Medulloblastoma. Nat Prod Bioprospect 2015 Mar 29. [Epub ahead of print].

15. Wei C, Xiao Q, Kuang X, Zhang T, Yang Z, Wang L. Fucoidan inhibits proliferation of the SKM-1 acute myeloid leukaemia cell line via the activation of apoptotic pathways and production of reactive oxygen species. Mol Med Rep 2015; 12:6649-6655.

16. Degagné E, Pandurangan A, Bandhuvula P, Kumar A, Eltanawy A, Zhang M, Yoshinaga Y, et al. Sphingosine-1 -phosphate lyase downregulation promotes colon carcinogenesis through STAT3-activated microRNAs. J Clin Invest 2014; 124:5368-5384.

17. Koskela HL, Eldfors S, Ellonen P, van Adrichem AJ, Kuusanmäki H, Andersson EI, et al. Somatic STAT3  mutations in large granular lymphocytic leukemia. N Engl J Med 2012; 366: 1905-1913.

18. Sun Y, Guo BF, Xu LB, Zhong JT, Liu ZW, Liang H, et al. STAT3-siRNA inhibits the growth of gastric cancer in vitro and in vivo. Cell Biochem Funct 2015; 33:495-502.

19. Xu Y, Zhang J, Wu J, Zhong S, Li H. Inhibition of JAK2 reverses paclitaxel resistance in human ovarian cancer cells. Int J Gynecol Cancer 2015; 25:1557-1564.

20. Miao L, Liu K, Xie M, Xing Y, Xi T. miR-375 inhibits Helicobacter pylori-induced gastric carcinogenesis by blocking JAK2-STAT3 signaling. Cancer Immunol Immunother 2014; 63: 699-711.

21. Um HJ, Min KJ, Kim DE, Kwon TK. Withaferin A inhibits JAK/STAT3 signaling and induces apoptosis of human renal carcinoma Caki cells. Biochem Biophys Res Commun 2012; 427:24 -29.

22. Li S, Priceman SJ, Xin H, Zhang W, Deng J, Liu Y, et al. Icaritin inhibits JAK/STAT3 signaling and growth of renal cell carcinoma. PLoS One 2013; 8:e81657.

23. Jeon YJ, Jung SN, Yun J, Lee CW, Choi J, Lee YJ, et al. Ginkgetin inhibits the growth of DU-145 prostate cancer cells through inhibition of signal transducer and activator of transcription 3 activity. Cancer Sci 2015; 106:413-420.

24. Xiong M, Wang L, Yu HL, Han H, Mao D, Chen J, et al. Ginkgetin exerts growth inhibitory and apoptotic effects on osteosarcoma cells through inhibition of STAT3 and activation of caspase-3/9.Oncol Rep 2016; 35:1034-1040.

25. Baek SH, Lee JH, Ko JH, Lee H, Nam D, Lee SG, et al. Ginkgetin blocks constitutive STAT3 activation and induces apoptosis through induction of SHP-1 and PTEN tyrosine phosphatases. Phytother Res 2016; 30:567-576.

26. Zhu S, Wang Z, Li Z, Peng H, Luo Y, Deng M, et al. Icaritin suppresses multiple myeloma, by inhibiting IL-6/JAK2/STAT3. Oncotarget 2015; 6:10460-10472.

27. Charles MS, Drunalini Perera PN, Doycheva DM, Tang J. Granulocyte-colony stimulating factor activates JAK2/PI3K/PDE3B pathway to inhibit corticosterone synthesis in a neonatal hypoxic-ischemic brain injury rat model. Exp Neurol 2015; 272:152-159. 

28. Guo H, German P, Bai S, Barnes S, Guo W, Qi X, et al. The PI3K/AKT Pathway and Renal Cell Carcinoma. J Genet Genomics 2015; 42:343-353.