Skip to main content

Advertisement

SpringerLink
Log in

Silencing Livin induces apoptotic and autophagic cell death, increasing chemotherapeutic sensitivity to cisplatin of renal carcinoma cells

  • Original Article
  • Published:
Tumor Biology

Abstract

Renal cell carcinoma (RCC) accounts for 3 % of all adult malignancies and is the most lethal urological cancer. Livin is a member of the inhibitor of apoptosis protein (IAP) family, which is associated with tumor resistance to radiotherapy and chemotherapy. Clinical data also showed that patients with high tumor grades and stages have higher expression levels of Livin in RCC cells. Autophagy is a survival mechanism activated in response to nutrient deprivation. A possible role of Livin in the autophagy of RCC cells has not been investigated; therefore, this pioneer study was carried out. Livin was silenced in RCC cells (slow virus infection [SVI]-shLivin cells) by lentiviral transfection. Then, mRNA and protein expression levels in the transfected cells were assessed by quantitative fluorescence PCR and Western blotting, respectively. In addition, acridine orange staining and electron microscopy were used to assess autophagy in SVI-shLivin cells. The cisplatin IC50 values for RCC cells were measured by the CCK8 assay. Potent antitumor activities were observed in xenograft mouse models generated with Livin-silenced RCC cells in terms of delayed tumor onset and suppressed tumor growth. These results suggested that Livin silencing could increase the chemotherapeutic sensitivity of RCC cells to cisplatin and induce autophagic cell death. A possible mechanism of Bcl-2 and Akt pathway involvement was discussed specifically in this study. Overall, Livin silencing induces apoptotic and autophagic cell death and increases chemotherapeutic sensitivity of RCC cells to cisplatin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Jemal A, Siegel R, Ward E, Murray T, Xu J. Smigal C and Thun M J. Cancer statistics, 2006. CA cancer j. Clin. 2006;56:106–30.

    Google Scholar 

  2. Flanigan RC, Campbell SC, Clark JI, Picken MM. Metastatic renal cell carcinoma. Curr Treat Options in Oncol. 2003;4:385–90.

    Article  Google Scholar 

  3. Yan B. Research progress on Livin protein: an inhibitor of apoptosis. Mol Cell Biochem. 2011;357(1–2):39–45.

    Article  CAS  PubMed  Google Scholar 

  4. Wang Y, Li Y, Zhou B, et al. Expression of the apoptosis inhibitor livin in colorectal adenoma-carcinoma sequence: correlations with pathology and outcome. Tumor Biol. 2014;35(12):11791–8.

    Article  CAS  Google Scholar 

  5. Wang J, Zhang X, Wei P, et al. Livin, Survivin and Caspase 3 as early recurrence markers in non-muscle-invasive bladder cancer. World J Urol. 2014;32(6):1477–84.

    Article  CAS  PubMed  Google Scholar 

  6. Ding ZY, Zhang H, Adell G, Olsson B, Sun XF. Livin expression is an independent factor in rectal cancer patients with or without preoperative radiotherapy. Radiat Oncol. 2013;8:281.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Zhao X, Yuan Y, Zhang Z, et al. Effects of shRNA-silenced livin and survivin on lung cancer cell proliferation and apoptosis. J BUON. 2014;19(3):757–62.

    PubMed  Google Scholar 

  8. Liu Y, Guo Q, Zhang H, et al. Effect of siRNA-livin on drug resistance to chemotherapy in glioma U251 cells and CD133 stem cells. Exp Ther Med. 2015;10(4):1317–23.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Chen N, Karantza-Wadsworth V. Role and regulation of autophagy in cancer. Biochim Biophys Acta. 2009;1793(9):1516–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Martin AP, Mitchell C, Rahmani M, Nephew KP, Grant S, Dent P. Inhibition of MCL-1 enhances lapatinib toxicity and overcomes lapatinib resistance via BAK-dependent autophagy. Cancer Biol Ther. 2009;8(21):2084–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Bonapace L, Bornhauser BC, Schmitz M, et al. Induction of autophagy-dependent necroptosis is required for childhood acute lymphoblastic leukemia cells to overcome glucocorticoid resistance. J Clin Invest. 2010;120(4):1310–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Turcotte S, Chan DA, Sutphin PD, Hay MP, Denny WA, Giaccia AJA. Molecule targeting VHL-deficient renal cell carcinoma that induces autophagy. Cancer Cell. 2008;8; 14(1):90–102.

    Article  Google Scholar 

  13. Lee JG, Shin JH, Shim HS, et al. Autophagy contributes to the chemo-resistance of non-small cell lung cancer in hypoxic conditions. Respir Res. 2015;16:138.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Sun WL, Chen J, Wang YP, Zheng H. Autophagy protects breast cancer cells from epirubicin-induced apoptosis and facilitates epirubicin-resistance development. Autophagy. 2011;7(9):1035–44.

    Article  CAS  PubMed  Google Scholar 

  15. Lv L, Liu HG, Dong SY, et al. Upregulation of CD44v6 contributes to acquired chemoresistance via the modulation of autophagy in colon cancer SW480 cells. Tumour Biol, 2016.

  16. Xu R, Liu S, Chen H, Lao L. MicroRNA-30a downregulation contributes to chemoresistance of osteosarcoma cells through activating Beclin-1-mediated autophagy. Oncol Rep. 2015.

  17. Yoon TM, Kim SA, Lee DH, et al. Expression of livin and the inhibition of tumor progression by livin silencing in laryngohypopharyngeal cancer. In Vivo. 2014;28(5):751–9.

    CAS  PubMed  Google Scholar 

  18. Song YH, Liao R, Li PC, Ge BO, Jiang LM, Gao LI, Zhang TY. Effects of siRNA livin on EJ human bladder cancer cells treated with mitomycin-C. Oncol Lett. 2015;10(4):2422–6.

    PubMed  PubMed Central  Google Scholar 

  19. Liu F, Chang H, Xu W, Zhai Y. The effects of livin shRNA on the response to cisplatin in HepG2 cells. Oncol Lett. 2015;10(5):2957–61.

    PubMed  PubMed Central  Google Scholar 

  20. Gu J, Ren L, Wang X, Qu C, Zhang Y. Expression of livin, survivin and caspase-3 in prostatic cancer and their clinical significance. Int J Clin Exp Pathol. 2015;1; 8(11):14034–9.

    Google Scholar 

  21. Ibrahim L, Aladle D, Mansour A, Hammad A, Al Wakeel AA. Abd el-Hameed SA. Expression and prognostic significance of livin/BIRC7 in childhood acute lymphoblastic leukemia. Med Oncol. 2014;31(5):941.

    Article  PubMed  Google Scholar 

  22. Myung DS, Park YL, Chung CY, et al. Expression of livin in colorectal cancer and its relationship to tumor cell behavior and prognosis. PLoS One. 2013;8(9):e73262.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Chung CY, Park YL, Park YL, et al. Myung DS, Kim JS, Cho SB, Lee WS, Joo YE. Expression and prognostic significance of Livin in gastric cancer. Oncol Rep 2013 ; 30(5):2520–2528.

  24. Kitamura H, Honma I, Torigoe T, et al. Expression of livin in renal cell carcinoma and detection of anti-livin autoantibody in patients. Urology. 2007;70(1):38–42.

    Article  PubMed  Google Scholar 

  25. Chang NC, Nguyen M, Germain M, Shore GC. Antagonism of Beclin1-de-pendent autophagy by BCL-2 at the endoplasmic reticulum requires NAF-1. EMBO J. 2010;29(3):606–18.

    Article  CAS  PubMed  Google Scholar 

  26. Casalino-Matsuda SM, Nair A, Beitel GJ, et al. Hypercapnia inhibits autophagy and bacterial killing in human macrophages by increasing expression of Bcl-2 and Bcl-xL. J Immunol. 2015;194(11):5388–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Fan J, Liu Y, Yin J, et al. Oxygen-glucose-deprivation/reoxygenation-induced autophagic cell death depends on JNK-mediated phosphorylation of Bcl-2. Cell Physiol Biochem. 2016;38(3):1063–74.

    Article  CAS  PubMed  Google Scholar 

  28. Trenti A, Grumati P, Cusinato F, Orso G, Bonaldo P, Trevisi L. Cardiac glycoside ouabain induces autophagic cell death in non-small cell lung cancer cells via a JNK-dependent decrease of Bcl-2. Biochem Pharmacol. 2014;89(2):197–209.

    Article  CAS  PubMed  Google Scholar 

  29. Liu K, Shi Y, et al. CHOP mediates ASPP2-induced autophagic apoptosis in hepatoma cells by releasing Beclin-1 from Bcl-2 and inducing nuclear translocation of Bcl-2. Cell Death Dis. 2014;5:e1323.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Kim SY, Yoo SJ, Ronnett GV, Kim EK, Moon C. Odorant stimulation promotes survival of rodent olfactory receptor neurons via PI3K/Akt activation and Bcl-2 expression. Mol Cells. 2015;38(6):535–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Zhang L, Zhang W, Wang YF, et al. Dual induction of apoptotic and autophagic cell death by targeting survivin in head neck squamous cell carcinoma. Cell Death and Disease. 2015;6:e1771.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Tang TK, Chiu SC, Lin CW, et al. Induction of survivin inhibition, G2/M cell cycle arrest and autophagic on cell death in human malignant glioblastoma cells. Chin J Physiol. 2015;58(2):95–103.

    CAS  PubMed  Google Scholar 

  33. Chiou SK, Hoa N, Hodges A. Sulindac sulfide induces autophagic death in gastric epithelial cells via survivin down-regulation: a mechanism of NSAIDs-induced gastric injury. Biochem Pharmacol. 2011;81(11):1317–23.

    Article  CAS  PubMed  Google Scholar 

  34. Wang YF, Zhang W, He KF, et al. Induction of autophagy-dependent cell death by the survivin suppressant YM155 in salivary adenoid cystic carcinoma. Apoptosis. 2014;19(4):748–58.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Zhen-Yu. Ding, Gui-Hong Liu, Birgit Olsson, Xiao-Feng S. Upregulation of the antiapoptotic factor livin contributes to cisplatin resistance in colon cancer cells. Tumour Biol. 2013;34(2):683–93.

    Article  Google Scholar 

  36. Kempkensteffen C, Hinz S, Christoph F, et al. Expression of the apoptosis inhibitor livin in renal cell carcinomas: correlations with pathology and outcome. Tumour Biol. 2007;28(3):132–8.

    Article  CAS  PubMed  Google Scholar 

  37. Wang Y, Li Y, Zhou B, et al. Expression of the apoptosis inhibitor livin in colorectal adenoma-carcinoma sequence: correlations with pathology and outcome. Tumour Biol. 2014;35(12):11791–8.

    Article  CAS  PubMed  Google Scholar 

  38. Liu S, Xie F, Wang H, Liu Z, Liu X, Sun L, Niu Z. Ubenimex inhibits cell proliferation, migration and invasion in renal cell carcinoma: the effect is autophagy-associated. Oncol Rep. 2015;33(3):1372–80.

    PubMed  Google Scholar 

  39. Wang XQ, Niu ZH, Jia Y, et al. Ubenimex inhibits cell proliferation, migration and invasion by inhibiting the expression of APN and inducing autophagic cell death in prostate cancer cells. Oncol Rep. 2015. doi:10.3892/or. 2016.4611.

    Google Scholar 

  40. Hamacher-Brady A, Brady NR. Bax/Bak-dependent, Drp1-independent targeting of X-linked inhibitor of apoptosis protein (XIAP) into inner mitochondrial compartments counteracts Smac/DIABLO-dependent effector caspase activation. J Biol Chem. 2015;290(36):22005–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Metwalli AR, Khanbolooki S, Jinesh G, et al. Smac mimetic reverses resistance to TRAIL and chemotherapy in human urothelial cancer cells. Cancer Biology & Therapy. 2010;10(9):885–92.

    Article  CAS  Google Scholar 

  42. Tamer E. Fandy, Sharmila Shankar, Srivastava RK, Smac/DIABLO enhances the therapeutic potential of chemotherapeutic drugs and irradiation, and sensitizes TRAIL-resistant breast cancer cells. Mol Cancer. 2008;7:60.

    Article  Google Scholar 

  43. Zhuang L, Shen LD, Li K, et al. Inhibition of livin expression suppresses cell proliferation and enhances chemosensitivity to cisplatin in human lung adenocarcinoma cells. Mol Med Rep. 2015;12(1):547–52.

    CAS  PubMed  Google Scholar 

  44. Kang CD, Ahn BK, Jeong CS, et al. Downregulation of JNK/SAPK activity is associated with the cross-resistance to P-glycoprotein-unrelated drugs in multidrug-resistant FM3A/M cells overexpressing P-glycoprotein. Exp Cell Res. 2000;256(1):300–7.

    Article  CAS  PubMed  Google Scholar 

  45. Palumbo C, De Luca A, Rosato N, et al. C-Jun N-terminal kinase activation by nitrobenzoxadiazoles leads to late-stage autophagy inhibition. J Transl Med. 2016;14:37. doi:10.1186/s12967-016-0796-x.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Nguyen TV, Sleiman M, Moriarty T, Herrick WG, Peyton SR. Sorafenib resistance and JNK signaling in carcinoma during extracellular matrix stiffening. Biomaterials. 2014;Jul; 35(22):5749–59.

    Article  PubMed  Google Scholar 

  47. He Q et al. MicroRNA-181a suppresses salivary adenoid cystic carcinoma metastasis by targeting MAPK-Snai2 pathway. Biochim Biophys Acta. 2013;1830(11):5258–66.

    Article  CAS  PubMed  Google Scholar 

  48. Chang B et al. Deregulation of Bmi-1 is associated with enhanced migration, invasion and poor prognosis in salivary adenoid cystic carcinoma. Biochim Biophys Acta. 2014;1840(12):3285–91.

    Article  CAS  PubMed  Google Scholar 

  49. Xie J et al. The antitumor effect of tanshinone IIA on anti-proliferation and decreasing VEGF/VEGFR2 expression on the human non-small cell lung cancer A549 cell-line. Acta Pharm Sin B. 2015;5(6):554–63.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

Thank Master Candidate Yin Lei for Statistical Analysis. This work was supported by the Shandong Key Research and Development Project (no. 2015GSF118055, 2014GGH218036 and 2014GSF118134), Medicine and Healthcare Technology Development Project of Shandong Province (no. 2014WS0341 and 2014WS0353), Natural Science Foundation of Shandong Province (no. 2014ZRB14513 and 2014ZRB14081), and National Natural Science Foundation of China (no.81602227).

Author information

Authors and Affiliations

  1. Department of Urology, Shandong Provincial Hospital Affiliated to Shandong University, 324 Jingwu Street, Jinan, 250021, People’s Republic of China

    Zhiyang Wang, Shuai Liu, Kejia Ding, Sentai Ding, Jiaju Lu, Dexuan Gao, Tong Zhang & Dongbin Bi

  2. Department of Digestive Diseases, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, 250021, People’s Republic of China

    Chensheng Li

Authors
  1. Zhiyang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuai Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kejia Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sentai Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chensheng Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jiaju Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dexuan Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tong Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dongbin Bi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dongbin Bi.

Ethics declarations

Conflicts of interest

None

Additional information

Zhiyang Wang and Shuai Liu have contributed equally to this work

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, Z., Liu, S., Ding, K. et al. Silencing Livin induces apoptotic and autophagic cell death, increasing chemotherapeutic sensitivity to cisplatin of renal carcinoma cells. Tumor Biol. 37, 15133–15143 (2016). https://doi.org/10.1007/s13277-016-5395-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-016-5395-1

Keywords

Search

Navigation