Keratin 6, induced by chronic cisplatin exposure, confers chemoresistance in human gastric carcinoma cells

Lim,Sung‑Chul; Parajuli,Keshab   Raj; Han,Song   Iy

doi:10.3892/or.2019.7201

Keratin 6, induced by chronic cisplatin exposure, confers chemoresistance in human gastric carcinoma cells

Authors:
- Sung‑Chul Lim
- Keshab Raj Parajuli
- Song Iy Han
View Affiliations

Affiliations: Department of Pathology, College of Medicine, Chosun University, Gwangju 61501, Republic of Korea, Research Center for Resistant Cells, College of Medicine, Chosun University, Gwangju 61501, Republic of Korea
Published online on: June 18, 2019 https://doi.org/10.3892/or.2019.7201
Pages: 797-804

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

Currently, various types of keratins have been reported to be highly expressed in cancer cells and to be associated with a malignant phenotype. In the present study, it was found that expression levels of keratin 6 (K6), keratin 16 (K16), and keratin 17 (K17) were highly elevated in SNU601 cells resistant to cisplatin (SNU601‑cis2 and SNU601‑cis10), but not in the parental SNU601 cells as confirmed by quantitative PCR, immunoblotting, and immunofluorescence assays. K6 is a type II keratin and is known to form a keratin filament in conjugation with type I keratin, K16 or K17. Thus, we attempted to understand the role of the overexpression of K6/K16 or K6/K17 keratin filaments by regulating the expression of K6. Silencing of K6 by siRNA in SNU601‑cis2 cells promoted oxaliplatin‑induced apoptosis in the resistant cells as shown by increased apoptotic body formation, caspase‑8 and caspase‑3 cleavage, and cytochrome c release. In addition, induction of K6 levels in wild‑type SNU601 cells, by transfection with pCMV6‑K6A and pCMV6‑K6B overexpression vectors, resulted in decreased apoptosis in response to cisplatin and L‑OHP. Platinum drugs, such as oxaliplatin, were shown to induce the extrinsic apoptotic pathway by inducing lipid raft formation and death receptor recruitment into lipid rafts. However, in the resistant cells, the oxaliplatin‑triggered extrinsic apoptotic pathway appeared to be suppressed by decreased lipid raft formation, and recruitment of death receptor 5 and FADD into lipid rafts. Therefore, the increase in the levels of the K6 filament may be associated with the regulation of lipid raft formation and may contribute, at least in part, to resistance to anticancer drugs.

View Figures

View References

1	Komatsu M, Sumizawa T, Mutoh M, Chen ZS, Terada K, Furukawa T, Yang XL, Gao H, Miura N, Sugiyama T and Akiyama S: Copper-transporting P-type adenosine triphosphatase (ATP7B) is associated with cisplatin resistance. Cancer Res. 60:1312–1316. 2000.PubMed/NCBI
2	Cort A, Ozben T, Saso L, De Luca C and Korkina L: Redox control of multidrug resistance and its possible modulation by antioxidants. Oxid Med Cell Longev. 2016:42519122016. View Article : Google Scholar : PubMed/NCBI
3	Wang J, Zhou JY and Wu GS: ERK-dependent MKP-1-mediated cisplatin resistance in human ovarian cancer cells. Cancer Res. 67:11933–11941. 2007. View Article : Google Scholar : PubMed/NCBI
4	Islam SU, Shehzad A, Sonn JK and Lee YS: PRPF overexpression induces drug resistance through actin cytoskeleton rearrangement and epithelial-mesenchymal transition. Oncotarget. 8:56659–56671. 2017. View Article : Google Scholar : PubMed/NCBI
5	Foerster F, Braig S, Moser C, Kubisch R, Busse J, Wagner E, Schmoeckel E, Mayr D, Schmitt S, Huettel S, et al: Targeting the actin cytoskeleton: Selective antitumor action via trapping PKCvarepsilon. Cell Death Dis. 5:e13982014. View Article : Google Scholar : PubMed/NCBI
6	Mukhtar E, Adhami VM and Mukhtar H: Targeting microtubules by natural agents for cancer therapy. Mol Cancer Ther. 13:275–284. 2014. View Article : Google Scholar : PubMed/NCBI
7	Xi J, Zhu X, Feng Y, Huang N, Luo G, Mao Y, Han X, Tian W, Wang G, Han X, et al: Development of a novel class of tubulin inhibitors with promising anticancer activities. Mol Cancer Res. 11:856–864. 2013. View Article : Google Scholar : PubMed/NCBI
8	Moll R, Divo M and Langbein L: The human keratins: Biology and pathology. Histochem Cell Biol. 129:705–733. 2008. View Article : Google Scholar : PubMed/NCBI
9	Moll R, Franke WW, Schiller DL, Geiger B and Krepler R: The catalog of human cytokeratins: Patterns of expression in normal epithelia, tumors and cultured cells. Cell. 31:11–24. 1982. View Article : Google Scholar : PubMed/NCBI
10	Fuchs E: Keratins and the skin. Annu Rev Cell Dev Biol. 11:123–153. 1995. View Article : Google Scholar : PubMed/NCBI
11	Coulombe PA and Omary MB: ‘Hard’ and ‘soft’ principles defining the structure, function and regulation of keratin intermediate filaments. Curr Opin Cell Biol. 14:110–122. 2002. View Article : Google Scholar : PubMed/NCBI
12	Galvin S, Loomis C, Manabe M, Dhouailly D and Sun TT: The major pathways of keratinocyte differentiation as defined by keratin expression: An overview. Adv Dermatol. 4:277–300. 1989.PubMed/NCBI
13	Van Muijen GN, Warnaar SO and Ponec M: Differentiation-related changes of cytokeratin expression in cultured keratinocytes and in fetal, newborn, and adult epidermis. Exp Cell Res. 171:331–345. 1987. View Article : Google Scholar : PubMed/NCBI
14	Gilbert S, Loranger A and Marceau N: Keratins modulate c-Flip/extracellular signal-regulated kinase 1 and 2 antiapoptotic signaling in simple epithelial cells. Mol Cell Biol. 24:7072–7081. 2004. View Article : Google Scholar : PubMed/NCBI
15	Roux A, Loranger A, Lavoie JN and Marceau N: Keratin 8/18 regulation of insulin receptor signaling and trafficking in hepatocytes through a concerted phosphoinositide-dependent Akt and Rab5 modulation. FASEB J. 31:3555–3573. 2017. View Article : Google Scholar : PubMed/NCBI
16	Lahdeniemi IAK, Misiorek JO, Antila CJM, Landor SK, Stenvall CA, Fortelius LE, Bergström LK, Sahlgren C and Toivola DM: Keratins regulate colonic epithelial cell differentiation through the Notch1 signalling pathway. Cell Death Differ. 24:984–996. 2017. View Article : Google Scholar : PubMed/NCBI
17	Srivastava SS, Alam H, Patil SJ, Shrinivasan R, Raikundalia S, Chaudhari PR and Vaidya MM: Keratin 5/14mediated cell differentiation and transformation are regulated by TAp63 and Notch1 in oral squamous cell carcinomaderived cells. Oncol Rep. 39:2393–2401. 2018.PubMed/NCBI
18	Chivu-Economescu M, Dragu DL, Necula LG, Matei L, Enciu AM, Bleotu C and Diaconu CC: Knockdown of KRT17 by siRNA induces antitumoral effects on gastric cancer cells. Gastric Cancer. 20:948–959. 2017. View Article : Google Scholar : PubMed/NCBI
19	Xu H, Choi SM, An CS, Min YD, Kim KC, Kim KJ and Choi CH: Concentration-dependent collateral sensitivity of cisplatin-resistant gastric cancer cell sublines. Biochem Biophys Res Commun. 328:618–622. 2005. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Iwasaki I, Sugiyama H, Kanazawa S and Hemmi H: Establishment of cisplatin-resistant variants of human neuroblastoma cell lines, TGW and GOTO, and their drug cross-resistance profiles. Cancer Chemother Pharmacol. 49:438–444. 2002. View Article : Google Scholar : PubMed/NCBI
22	Sun CL and Chao CC: Cross-resistance to death ligand-induced apoptosis in cisplatin-selected HeLa cells associated with overexpression of DDB2 and subsequent induction of cFLIP. Mol Pharmacol. 67:1307–1314. 2005. View Article : Google Scholar : PubMed/NCBI
23	Xu L, Qu X, Zhang Y, Hu X, Yang X, Hou K, Teng Y, Zhang J, Sada K and Liu Y: Oxaliplatin enhances TRAIL-induced apoptosis in gastric cancer cells by CBL-regulated death receptor redistribution in lipid rafts. FEBS Lett. 583:943–948. 2009. View Article : Google Scholar : PubMed/NCBI
24	Huang CR, Jin ZX, Dong L, Tong XP, Yue S, Kawanami T, Sawaki T, Sakai T, Miki M, Iwao H, et al: Cisplatin augments FAS-mediated apoptosis through lipid rafts. Anticancer Res. 30:2065–2071. 2010.PubMed/NCBI
25	Rothnagel JA, Seki T, Ogo M, Longley MA, Wojcik SM, Bundman DS, Bickenbach JR and Roop DR: The mouse keratin 6 isoforms are differentially expressed in the hair follicle, footpad, tongue and activated epidermis. Differentiation. 65:119–130. 1999. View Article : Google Scholar : PubMed/NCBI
26	Wong P and Coulombe PA: Loss of keratin 6 (K6) proteins reveals a function for intermediate filaments during wound repair. J Cell Biol. 163:327–337. 2003. View Article : Google Scholar : PubMed/NCBI
27	Jiang CK, Magnaldo T, Ohtsuki M, Freedberg IM, Bernerd F and Blumenberg M: Epidermal growth factor and transforming growth factor alpha specifically induce the activation- and hyperproliferation-associated keratins 6 and 16. Proc Natl Acad Sci USA. 90:6786–6790. 1993. View Article : Google Scholar : PubMed/NCBI
28	Leigh IM, Navsaria H, Purkis PE, McKay IA, Bowden PE and Riddle PN: Keratins (K16 and K17) as markers of keratinocyte hyperproliferation in psoriasis in vivo and in vitro. Br J Dermatol. 133:501–511. 1995. View Article : Google Scholar : PubMed/NCBI
29	Gilbert S, Loranger A, Daigle N and Marceau N: Simple epithelium keratins 8 and 18 provide resistance to Fas-mediated apoptosis. The protection occurs through a receptor-targeting modulation. J Cell Biol. 154:763–773. 2001. View Article : Google Scholar : PubMed/NCBI
30	Bauman PA, Dalton WS, Anderson JM and Cress AE: Expression of cytokeratin confers multiple drug resistance. Proc Natl Acad Sci USA. 91:5311–5314. 1994. View Article : Google Scholar : PubMed/NCBI
31	Anderson JM, Heindl LM, Bauman PA, Ludi CW, Dalton WS and Cress AE: Cytokeratin expression results in a drug-resistant phenotype to six different chemotherapeutic agents. Clin Cancer Res. 2:97–105. 1996.PubMed/NCBI
32	Wang Y, He QY, Tsao SW, Cheung YH, Wong A and Chiu JF: Cytokeratin 8 silencing in human nasopharyngeal carcinoma cells leads to cisplatin sensitization. Cancer Lett. 265:188–196. 2008. View Article : Google Scholar : PubMed/NCBI
33	Ricciardelli C, Lokman NA, Pyragius CE, Ween MP, Macpherson AM, Ruszkiewicz A, Hoffmann P and Oehler MK: Keratin 5 overexpression is associated with serous ovarian cancer recurrence and chemotherapy resistance. Oncotarget. 8:17819–17832. 2017. View Article : Google Scholar : PubMed/NCBI
34	Gilbert S, Loranger A, Lavoie JN and Marceau N: Cytoskeleton keratin regulation of FasR signaling through modulation of actin/ezrin interplay at lipid rafts in hepatocytes. Apoptosis. 17:880–894. 2012. View Article : Google Scholar : PubMed/NCBI
35	Gilbert S, Loranger A, Omary MB and Marceau N: Keratin impact on PKCδ- and ASMase-mediated regulation of hepatocyte lipid raft size-implication for FasR-associated apoptosis. J Cell Sci. 129:3262–3273. 2016. View Article : Google Scholar : PubMed/NCBI