Proteasome inhibitor lactacystin enhances cisplatin cytotoxicity by increasing endoplasmic reticulum stress‑associated apoptosis in HeLa cells

Xu,Ye; Li,Di; Zeng,Linchuan; Wang,Chunyan; Zhang,Lili; Wang,Yan; Yu,Yang; Liu,Shibing; Li,Zhixin

doi:10.3892/mmr.2014.2683

Proteasome inhibitor lactacystin enhances cisplatin cytotoxicity by increasing endoplasmic reticulum stress‑associated apoptosis in HeLa cells

Authors:
- Ye Xu
- Di Li
- Linchuan Zeng
- Chunyan Wang
- Lili Zhang
- Yan Wang
- Yang Yu
- Shibing Liu
- Zhixin Li
View Affiliations

Affiliations: Medical Research Laboratory, Jilin Medical College, Jilin, Jilin 132013, P.R. China, Department of Urology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Histology and Embryology, Jilin Medical College, Jilin, Jilin 132013, P.R. China
Published online on: October 16, 2014 https://doi.org/10.3892/mmr.2014.2683
Pages: 189-195
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cisplatin is commonly used as a therapeutic agent, despite its known adverse side effects and the occurrence of drug resistance. The development of novel methods for combination therapy with cisplatin is required in order to circumvent these limitations of cisplatin alone. The proteasome inhibitor lactacystin (LAC) has been indicated to produce anti‑tumor effects, and has previously been used as an antitumor agent in cancer treatment research; however, its effects in combination with cisplatin treatment are unknown. In the current study, the effects of LAC in combination with cisplatin treatment were investigated in HeLa human cervical cancer (HCC) cells. The results demonstrated that cisplatin treatment inhibited cell growth and induced cell apoptosis. HeLa cell exposure to cisplatin induced endoplasmic reticulum (ER) stress‑associated apoptosis, and LAC treatment increased levels of cell apoptosis and the activation of caspase‑3. Specifically, LAC treatment increased the cisplatin‑induced expression of PDI, GRP78, CHOP, cleaved caspase‑4 and cleaved caspase‑3. Together, these data indicate that LAC is able to enhance cisplatin cytotoxicity by increasing ER stress‑associated apoptosis in HeLa cells.

View Figures

View References

1	Muggia F: Platinum compounds 30 years after the introduction of cisplatin: implications for the treatment of ovarian cancer. Gynecol Oncol. 112:275–281. 2009.PubMed/NCBI
2	Florea AM and Büsselberg D: Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects. Cancers (Basel). 3:1351–1371. 2011. View Article : Google Scholar : PubMed/NCBI
3	Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, Castedo M and Kroemer G: Molecular mechanisms of cisplatin resistance. Oncogene. 31:1869–1883. 2012. View Article : Google Scholar
4	Eastman A: The formation, isolation and characterization of DNA adducts produced by anticancer platinum complexes. Pharmacol Ther. 34:155–166. 1987. View Article : Google Scholar : PubMed/NCBI
5	Woźniak K and Błasiak J: Recognition and repair of DNA-cisplatin adducts. Acta Biochim Pol. 49:583–596. 2002.
6	Basu A and Krishnamurthy S: Cellular responses to Cisplatin-induced DNA damage. J Nucleic Acids. 2010:pii: 201367. 2010. View Article : Google Scholar
7	Mandic A, Hansson J, Linder S and Shoshan MC: Cisplatin induces endoplasmic reticulum stress and nucleus-independent apoptotic signaling. J Biol Chem. 278:9100–9106. 2003. View Article : Google Scholar : PubMed/NCBI
8	Liu H and Baliga R: Endoplasmic reticulum stress-associated caspase 12 mediates cisplatin-induced LLC-PK1 cell apoptosis. J Am Soc Nephrol. 16:1985–1992. 2005. View Article : Google Scholar : PubMed/NCBI
9	Peyrou M, Hanna PE and Cribb AE: Cisplatin, gentamicin, and p-aminophenol induce markers of endoplasmic reticulum stress in the rat kidneys. Toxicol Sci. 99:346–353. 2007. View Article : Google Scholar : PubMed/NCBI
10	Yu F, Megyesi J and Price PM: Cytoplasmic initiation of cisplatin cytotoxicity. Am J Physiol Renal Physiol. 295:F44–F52. 2008. View Article : Google Scholar : PubMed/NCBI
11	Berridge MJ: The endoplasmic reticulum: a multifunctional signaling organelle. Cell Calcium. 32:235–249. 2002. View Article : Google Scholar : PubMed/NCBI
12	Jørgensen MM, Bross P and Gregersen N: Protein quality control in the endoplasmic reticulum. APMIS Suppl. 109:86–91. 2003.
13	Rao RV, Ellerby HM and Bredesen DE: Coupling endoplasmic reticulum stress to the cell death program. Cell Death Differ. 11:372–380. 2004. View Article : Google Scholar : PubMed/NCBI
14	Kim R, Emi M, Tanabe K and Murakami S: Role of the unfolded protein response in cell death. Apoptosis. 11:5–13. 2006. View Article : Google Scholar : PubMed/NCBI
15	Benbrook DM and Long A: Integration of autophagy, proteasomal degradation, unfolded protein response and apoptosis. Exp Oncol. 34:286–297. 2012.PubMed/NCBI
16	Høyer-Hansen M and Jäättelä M: Connecting endoplasmic reticulum stress to autophagy by unfolded protein response and calcium. Cell Death Differ. 14:1576–1582. 2007.PubMed/NCBI
17	Yu H, Su J, Xu Y, Kang J, Li H, Zhang L, Yi H, Xiang X, Liu F and Sun L: p62/SQSTM1 involved in cisplatin resistance in human ovarian cancer cells by clearing ubiquitinated proteins. Eur J Cancer. 47:1585–1594. 2011. View Article : Google Scholar : PubMed/NCBI
18	Xu Y, Yu H, Qin H, Kang J, et al: Inhibition of autophagy enhances cisplatin cytotoxicity through endoplasmic reticulum stress in human cervical cancer cells. Cancer Lett. 314:232–243. 2012. View Article : Google Scholar : PubMed/NCBI
19	Zhong JT, Xu Y, Yi HW, et al: The BH3 mimetic S1 induces autophagy through ER stress and disruption of Bcl-2/Beclin 1 interaction in human glioma U251 cells. Cancer Lett. 323:180–187. 2012. View Article : Google Scholar : PubMed/NCBI
20	Liu N, Xu Y, Sun JT, et al: The BH3 mimetic S1 induces endoplasmic reticulum stress-associated apoptosis in cisplatin-resistant human ovarian cancer cells although it activates autophagy. Oncol Rep. 30:2677–2684. 2013.
21	Fenteany G, Standaert RF, Lane WS, Choi S, Corey EJ and Schreiber SL: Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin. Science. 268:726–731. 1995. View Article : Google Scholar : PubMed/NCBI
22	Craiu A, Gaczynska M, Akopian T, et al: Lactacystin and clasto-lactacystin beta-lactone modify multiple proteasome beta-subunits and inhibit intracellular protein degradation and major histocompatibility complex class I antigen presentation. J Biol Chem. 272:13437–13445. 1997. View Article : Google Scholar
23	Ostrowska H, Wojcik C, Omura S and Worowski K: Lactacystin, a specific inhibitor of the proteasome, inhibits human platelet lysosomal cathepsin A-like enzyme. Biochem Biophys Res Commun. 234:729–732. 1997. View Article : Google Scholar : PubMed/NCBI
24	Geier E, Pfeifer G, Wilm M, et al: A giant protease with potential to substitute for some functions of the proteasome. Science. 283:978–981. 1999. View Article : Google Scholar : PubMed/NCBI
25	Rebillard A, Lagadic-Gossmann D and Dimanche-Boitrel MT: Cisplatin cytotoxicity: DNA and plasma membrane targets. Curr Med Chem. 15:2656–2663. 2008. View Article : Google Scholar : PubMed/NCBI
26	Ogata M, Hino S, Saito A, Morikawa K, et al: Autophagy is activated for cell survival after endoplasmic reticulum stress. Mol Cell Biol. 26:9220–9231. 2006. View Article : Google Scholar : PubMed/NCBI
27	Lin X, Li S, Zhao Y, Ma X, Zhang K, He X and Wang Z: Interaction domains of p62: a bridge between p62 and selective autophagy. DNA Cell Biol. 32:220–227. 2013. View Article : Google Scholar : PubMed/NCBI
28	Tanida I: Autophagosome formation and molecular mechanism of autophagy. Antioxid Redox Signal. 14:2201–2214. 2011. View Article : Google Scholar : PubMed/NCBI
29	Szegezdi E, Logue SE, Gorman AM and Samali A: Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep. 7:880–885. 2006. View Article : Google Scholar : PubMed/NCBI
30	Tabas I and Ron D: Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress. Nat Cell Biol. 13:184–190. 2011. View Article : Google Scholar : PubMed/NCBI
31	Olive PL and Banáth JP: Kinetics of H₂AX phosphorylation after exposure to cisplatin. Cytometry B Clin Cytom. 76:79–90. 2009.