Simultaneous inhibition of atypical protein kinase‑C and mTOR impedes bladder cancer cell progression

Patel,Rekha; Islam,SM Anisul; Bommareddy,Raja  Reddy; Smalley,Tracess; Acevedo‑Duncan,Mildred

doi:10.3892/ijo.2020.5021

Simultaneous inhibition of atypical protein kinase‑C and mTOR impedes bladder cancer cell progression

Authors:
- Rekha Patel
- SM Anisul Islam
- Raja Reddy Bommareddy
- Tracess Smalley
- Mildred Acevedo‑Duncan
View Affiliations

Affiliations: Department of Chemistry, University of South Florida, Tampa, FL 33620, USA
Published online on: March 20, 2020 https://doi.org/10.3892/ijo.2020.5021
Pages: 1373-1386
Copyright: © Patel et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Despite enormous scientific advancements in cancer treatment, there is a need for research to combat cancer, particularly bladder cancer. Drugs once proved to be effective in treating bladder cancer have shown reduced efficacy; hence, the cancer recurrence rate is increasing. To overcome this situation, several strategies have been considered, including the development of novel active drugs or modification of existing therapeutic regimens by combining two or more existing drugs. In recent years, atypical protein kinase Cs (PKCs), phospholipid‑dependent serine/threonine kinases, have been considered as a central regulator of various cancer‑associated signaling pathways, and they control cell cycle progression, tumorigenesis and metastasis. Additionally, the biologically crucial mTOR signaling pathway is altered in numerous types of cancer, including bladder cancer. Furthermore, despite independent activation, atypical PKC signaling can be triggered by mTOR. The present study examined whether the concurrent inhibition of atypical PKCs and mTOR using a combination of novel atypical PKC inhibitors (ICA‑I, an inhibitor of PKC‑ι; or ζ‑Stat, an inhibitor of PKC‑ζ) and rapamycin blocks bladder cancer progression. In the present study, healthy bladder MC‑SV‑HUCT2 and bladder cancer TCCSUP cells were tested and subjected to a WST1 assay, western blot analysis, immunoprecipitation, a scratch wound healing assay, flow cytometry and immunofluorescence analyses. The results revealed that the combination therapy induced a reduction in human bladder cancer cell viability compared with control and individual atypical PKC inhibitor and rapamycin treatment. Additionally, the concurrent inhibition of atypical PKCs and mTOR retards the migration of bladder cancer cells. These findings indicated that the administration of atypical PKC inhibitors together with rapamycin could be a useful therapeutic option in treating bladder cancer.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2019. CA Cancer J Clin. 69:7–34. 2019. View Article : Google Scholar : PubMed/NCBI
2	Montironi R, Cheng L, Scarpelli M and Lopez-Beltran A: Pathology and genetics: Tumours of the urinary system and male genital system: Clinical implications of the 4th edition of the WHO classification and beyond. Eur Urol. 70:120–123. 2016. View Article : Google Scholar : PubMed/NCBI
3	Abraham S, Knapp DW, Cheng L, Snyder PW, Mittal SK, Bangari DS, Kinch M, Wu L, Dhariwal J and Mohammed SI: Expression of EphA2 and Ephrin A-1 in carcinoma of the urinary bladder. Clin Cancer Res. 12:353–360. 2006. View Article : Google Scholar : PubMed/NCBI
4	Reuter VE: Pathology of bladder cancer: Assessment of prognostic variables and response to therapy. Semin Oncol. 17:524–532. 1990.PubMed/NCBI
5	Sylvester RJ, van der MEIJDEN AP and Lamm DL: Intravesical bacillus Calmette-Guerin reduces the risk of progression in patients with superficial bladder cancer: A meta-analysis of the published results of randomized clinical trials. J Urol. 168:1964–1970. 2002. View Article : Google Scholar : PubMed/NCBI
6	Gschwend JE, Dahm P and Fair WR: Disease specific survival as endpoint of outcome for bladder cancer patients following radical cystectomy. Eur Urol. 41:440–448. 2002. View Article : Google Scholar : PubMed/NCBI
7	Nelson DL and Cox MM: Lehninger principles of biochemistry. 5th ed. Book; pp. 1–1294. 2008
8	Islam SMA, Patel R and Acevedo-Duncan M: Protein Kinase C-ζ stimulates colorectal cancer cell carcinogenesis via PKC-ζ/Rac1/Pak1/β-Catenin signaling cascade. Biochim Biophys Acta Mol cell Res. 1865:650–664. 2018. View Article : Google Scholar : PubMed/NCBI
9	Islam SMA, Patel R, Bommareddy RR, Khalid KM and Acevedo-Duncan M: The modulation of actin dynamics via atypical Protein Kinase-C activated Cofilin regulates metastasis of colorectal cancer cells. Cell Adh Migr. 13:106–120. 2019. View Article : Google Scholar :
10	Shaw RJ and Cantley LC: Ras, PI(3)K and mTOR signalling controls tumour cell growth. Nature. 441:424–430. 2006. View Article : Google Scholar : PubMed/NCBI
11	Liu P, Cheng H, Roberts TM and Zhao JJ: Targeting the phos-phoinositide 3-kinase pathway in cancer. Nat Rev Drug Discov. 8:627–644. 2009. View Article : Google Scholar : PubMed/NCBI
12	Sengupta S, Peterson TR and Sabatini DM: Regulation of the mTOR complex 1 pathway by nutrients, growth factors, and stress. Mol Cell. 40:310–322. 2010. View Article : Google Scholar : PubMed/NCBI
13	Laplante M and Sabatini DM: mTOR signaling in growth control and disease. Cell. 149:274–293. 2012. View Article : Google Scholar : PubMed/NCBI
14	Facchinetti V, Ouyang W, Wei H, Soto N, Lazorchak A, Gould C, Lowry C, Newton AC, Mao Y, Miao RQ, et al: The mammalian target of rapamycin complex 2 controls folding and stability of Akt and protein kinase C. EMBO J. 27:1932–1943. 2008. View Article : Google Scholar : PubMed/NCBI
15	Ikenoue T, Inoki K, Yang Q, Zhou X and Guan KL: Essential function of TORC2 in PKC and Akt turn motif phosphorylation, maturation and signalling. EMBO J. 27:1919–1931. 2008. View Article : Google Scholar : PubMed/NCBI
16	Sarbassov DD, Guertin DA, Ali SM and Sabatini DM: Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science. 307:1098–1101. 2005. View Article : Google Scholar : PubMed/NCBI
17	Eder AM, Sui X, Rosen DG, Nolden LK, Cheng KW, Lahad JP, Kango-Singh M, Lu KH, Warneke CL, Atkinson EN, et al: Atypical PKC contributes to poor prognosis through loss of apical-basal polarity and Cyclin E overexpression in ovarian cancer. Proc Natl Acad Sci. 102:12519–12524. 2005. View Article : Google Scholar
18	Guo H, Ma Y, Zhang B, Sun B, Niu R, Ying G and Zhang N: Pivotal Advance: PKC is required for migration of macrophages. J Leukoc Biol. 85:911–918. 2009. View Article : Google Scholar : PubMed/NCBI
19	Smalley T, Islam SMA, Apostolatos C, Apostolatos A and Acevedo-Duncan M: Analysis of PKC-ζ protein levels in normal and malignant breast tissue subtypes. Oncol Lett. 17:1537–1546. 2019.PubMed/NCBI
20	Islam SMA, Dey A and Acevedo-Duncan M: Abstract 719: Combination of atypical protein kinase-C inhibitor and 5-fluo-rouracil retards the proliferation of colorectal cancer cells. Mol Cell Biol. 719:2019.
21	Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
22	Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72:248–254. 1976. View Article : Google Scholar : PubMed/NCBI
23	Patel R, Win H, Desai S, Patel K, Matthews JA and Acevedo-Duncan M: Involvement of PKC-ι in glioma proliferation. Cell Prolif. 41:122–135. 2008. View Article : Google Scholar : PubMed/NCBI
24	Sherr CJ and Roberts JM: Living with or without cyclins and cyclin-dependent kinases. Genes Dev. 18:2699–2711. 2004. View Article : Google Scholar : PubMed/NCBI
25	Hardie DG and Hanks S: The protein kinase factsbook. Academic Press; 1995
26	Kriegmair MC, Balk M, Wirtz R, Steidler A, Weis CA, Breyer J, Hartmann A, Bolenz C and Erben P: Expression of the p53 inhibitors mdm2 and mdm4 as outcome predictor in muscle-invasive bladder cancer. Anticancer Res. 36:5205–5213. 2016. View Article : Google Scholar : PubMed/NCBI
27	Wang AS, Ong PF, Chojn owski A, Clavel C and Dreesen O: Loss of lamin B1 is a biomarker to quantify cellular senescence in photoaged skin. Sci Rep. 7:156782017. View Article : Google Scholar : PubMed/NCBI
28	Goldstein S: Replicative senescence: The human fibroblast comes of age. Science. 249:1129–1133. 1990. View Article : Google Scholar : PubMed/NCBI
29	Wheelock MJ and Johnson KR: Cadherins as Modulators of Cellular Phenotype. Annu Rev Cell Dev Biol. 19:207–235. 2003. View Article : Google Scholar : PubMed/NCBI
30	De P, Carlson JH, Wu H, Marcus A, Leyland-Jones B and Dey N: Wnt-beta-catenin pathway signals metastasis-associated tumor cell phenotypes in triple negative breast cancers. Oncotarget. 7:43124–43149. 2016. View Article : Google Scholar : PubMed/NCBI
31	Mauro L, Sisci D, Bartucci M, Salerno M, Kim J, Tam T, Guvakova MA, Ando S and Surmacz E: SHC-α5β1 integrin interactions regulate breast cancer cell adhesion and motility. Exp Cell Res. 252:439–448. 1999. View Article : Google Scholar : PubMed/NCBI
32	Jackson JG, Yoneda T, Clark GM and Yee D: Elevated levels of p66 Shc are found in breast cancer cell lines and primary tumors with high metastatic potential. Clin Cancer Res. 6:1135–1139. 2000.PubMed/NCBI
33	Xiao L, Wang YC, Li WS and Du Y: The role of mTOR and phospho-p70S6K in pathogenesis and progression of gastric carcinomas: An immunohistochemical study on tissue micro-array. J Exp Clin Cancer Res. 28:1522009. View Article : Google Scholar
34	Malley CO and Pidgeon GP: The mTOR pathway in obesity driven gastrointestinal cancers: Potential targets and clinical trials. BBA Clin. 5:29–40. 2016. View Article : Google Scholar : PubMed/NCBI
35	Ching CB and Hansel DE: Expanding therapeutic targets in bladder cancer: The PI3K/Akt/mTOR pathway. Lab Invest. 90:1406–1414. 2010. View Article : Google Scholar : PubMed/NCBI
36	Carneiro BA, Meeks JJ, Kuzel TM, Scaranti M, Abdulkadir SA and Giles FJ: Emerging therapeutic targets in bladder cancer. Cancer Treat Rev. 41:170–178. 2015. View Article : Google Scholar
37	Apostolatos AH, Ratnayake WS, Win-Piazza H, Apostolatos CA, Smalley T, Kang L, Salup R, Hill R and Acevedo-Duncan M: Inhibition of atypical protein kinase C-ι effectively reduces the malignancy of prostate cancer cells by downregulating the NF-κB signaling cascade. Int J Oncol. 53:1836–1846. 2018.PubMed/NCBI
38	Ratnayake WS, Apostolatos AH, Ostrov DA and Acevedo-Duncan M: Two novel atypical PKC inhibitors; ACPD and DNDA effectively mitigate cell proliferation and epithelial to mesenchymal transition of metastatic melanoma while inducing apoptosis. Int J Oncol. 51:1370–1382. 2017. View Article : Google Scholar : PubMed/NCBI
39	Pinto-Leite R, Arantes-Rodrigues R, Sousa N, Oliveira PA and Santos L: mTOR inhibitors in urinary bladder cancer. Tumor Biol. 37:11541–11551. 2016. View Article : Google Scholar
40	Alayev A, Salamon RS, Schwartz NS, Berman AY, Wiener SL and Holz MK: Combination of rapamycin and resveratrol for treatment of bladder cancer. J Cell Physiol. 232:436–446. 2017. View Article : Google Scholar
41	Alayev A, Berger SM, Kramer MY, Schwartz NS and Holz MK: The combination of rapamycin and resveratrol blocks autophagy and induces apoptosis in breast cancer cells. J Cell Biochem. 116:450–547. 2015. View Article : Google Scholar :
42	Islam SMA, Dey A, Patel R, Smalley T and Acevedo-Duncan M: Atypical Protein Kinase-C inhibitors exhibit a synergistic effect in facilitating DNA damaging effect of 5-fluorouracil in colorectal cancer cells. Biomed Pharmacother. 120:1096652020. View Article : Google Scholar
43	Orlando DA, Lin CY, Bernard A, Wang JY, Socolar JE, Iversen ES, Hartemink AJ and Haase SB: Global control of cell-cycle transcription by coupled CDK and network oscillators. Nature. 453:944–947. 2008. View Article : Google Scholar : PubMed/NCBI
44	Levin VA, Panchabhai SC, Shen L, Kornblau SM, Qiu Y and Baggerly KA: Different changes in protein and phosphoprotein levels result from serum starvation of high-grade glioma and adenocarcinoma cell lines. J Proteome Res. 9:179–191. 2010. View Article : Google Scholar
45	Ballou LM and Lin RZ: Rapamycin and mTOR kinase inhibitors. J Chem Biol. 1:27–36. 2008. View Article : Google Scholar