Microarray based gene expression profiling of advanced gall bladder cancer
DOI:
https://doi.org/10.32471/exp-oncology.2312-8852.vol-42-no-4.15476Keywords:
Cdc45, chronic cholecystitis, gall bladder cancer, MCM4, microarrayAbstract
Summary. Background: Gall bladder cancer (GBC) is an aggressive cancer with specific predilection like female gender and specific geographical areas, however the molecular mechanisms and factors contributing to the clinical or biological behavior are not understood. Aim: The aim of this study was to perform a comprehensive analysis of differentially expressed genes in advanced GBC and chronic cholecystitis (CC) cases. Materials and Methods: Microarray was planned on fresh specimens of advanced GBC and CC cases using single color cRNA based microarray technique (8X60K format; Agilent Technologies, USA). Twelve advanced GBC and four CC patients were included in the study. Results: Of the total of 1307 differentially expressed genes, 535 genes were significantly upregulated, while 772 genes were significantly downregulated in advanced GBC vs CC samples. Differentially expressed genes were associated with biological processes (55.03%), cellular components (31.48%), and molecular functions (13.49%) respectively. The important pathways or key processes affected were cell cycle, DNA replication, oxidative stress, gastric cancer pathway. Using in silico analysis tools, three differentially expressed genes i.e. TPX2, Cdc45 and MCM4 were selected (for their significant role in DNA replication and microtubule function) and were further validated in 20 advanced GBC cohort by immunohistochemistry. Significant positive association of Cdc45 and MCM4 proteins was found in advanced GBC cases (p = 0.043), suggesting the probable oncogenic role of Cdc45 and MCM4 proteins in advanced GBC. Conclusion: Our data demonstrate the potential regulation of Cdc45-MCM4 axis in advanced GBC tumors. Additionally, our study also revealed a range of differentially expressed genes (e.g. TPX2, AKURA etc.) between GBC and CC, and further validation of these genes might provide a potential diagnostic or therapeutic target in future.
References
Bizama C, Garcia P, Espinoza JA, et al. Targeting specific molecular pathways holds promise for advanced gallbladder cancertherapy. Cancer Treat Rev 2015; 41: 222–34.
Randi G, Franceschi S, La Vecchia C. Gallbladder cancer worldwide: geographical distribution and risk factors. Int J Cancer 2006; 118: 1591–602.
Kumari N, Corless CL, Warrick A, et al. Mutation profiling in gallbladder cancer in Indian population. Indian J Pathol Microbiol 2014; 57: 9–12.
Qu K, Zhang X, Cui R, et al. Meta-signature of mutated genes in gallbladder cancer: evidence based high throughput screening assays. Ann Transl Med 2016; 4: 229.
Kim JH, Kim HN, Lee KT et al. Gene expression profiles in gallbladder cancer: the close genetic similarity seen for early and advanced gallbladder cancers may explain the poor prognosis. Tumor Biol 2008; 29: 41–9.
Li SH, Li CF, Sung MT, et al. SKP2 is an independent prognosticator of gallbladder carcinoma among p27(Kip1)-interacting cell cycle regulators: an immunohistochemical study of 62 cases by tissue microarray. Mod Pathol 2007; 20: 497–507.
Ghosh M, Sakhuja P, Singh S, et al. p53 and beta-catenin expression in gallbladder tissues and correlation with tumor progression in gallbladder cancer. Saudi J Gastroenterol 2013; 19: 34–9.
Gupta V, Goel MM, Chandra A, et al. Expression and clinicopathological significance of antiapoptotis protein survivin in gallbladder cancer. Indian J Pathol Microbiol. 2016; 59: 143–7.
Espinoza JA, Riquelme I, Sagredo EA, et al. Mucin 5B, carbonic anhydrase 9 and claudin 18 are potential theranostic markers of gallbladder carcinoma. Histopathology 2019; 74: 597–607.
Sharma A, Dwary AD, Mohanti BK, et al. Best supportive care compared with chemotherapy for unresectable gall bladder cancer: a randomized controlled study. J Clin Oncol 2010; 28: 4581–6.
Chu L, Scharf E, Kondo T. GeneSpring TM: Tools for analyzing microarray expression data. Genome Info 2001; 12: 227–9.
Ashburner M, Ball CA, Blake JA, et al. Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet 2000; 25: 25–9.
Wong AK, Krishnan A, Troyanskaya OG. GIANT 2.0: genome-scale integrated analysis of gene networks in tissues. Nucl Acid Res 2018; 46: 65–70.
Engeland K. Cell cycle arrest through indirect transcriptional repression by p53: I have a DREAM. Cell Death Diff 2018; 25: 114–32.
Batra Y, Pal S, Dutta U, et al. Gallbladder cancer in India: a dismal picture. J Gastroenterol Hepatol 2005; 20: 309–14.
Hundal R, Shaffer EA. Gallbladder cancer: epidemiology and outcome. Clin Epidem 2014; 6: 99–109.
Stancu M, Cгruntu ID, Sajin M, et al. Immunohistochemical markers in the study of gallbladder premalignant lesions and cancer. Rev Med Chir Soc Med Nat Iasi 2007; 111: 734–43.
Agrawal V, Goel A, Krishnani N, et al. p53, carcinoembryonic antigen and carbohydrate antigen 19.9 expression in gall bladder cancer, precursor epithelial lesions and xanthogranulomatous cholecystitis. J Postgrad Med 2010; 56: 262–6.
Ghosh M, Sakhuja P, Singh S, et al. p53 and beta-catenin expression in gallbladder tissues and correlation with tumor progression in gallbladder cancer. Saudi J Gastroenterol 2013; 19: 34–9.
Gupta V, Goel MM, Chandra A, et al. Expression and clinicopathological significance of antiapoptotis protein survivin in gallbladder cancer. Indian J Pathol Microbiol 2016; 59: 143–7.
Mikami T, Yanagisawa N, Baba H, et al. Association of Bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis. Cancer 1999; 85: 318–25.
Yang D, Zhan M, Chen T, et al. miR-125b-5p enhances chemotherapy sensitivity to cisplatin by down-regulating Bcl2 in gallbladder cancer. Sci Rep 2017; 7: 43109.
Nigam J, Chandra A, Kazmi HR, et al. Prognostic significance of survivin in resected gallbladder cancer. J Surg Res 2015; 194: 57–62.
Xiang S, Wang Z, Ye Y, et al. E2F1 and E2F7 differentially regulate KPNA2 to promote the development of gallbladder cancer. Oncogene 2019; 38: 1269–81.
Li M, Zhang F, Wang X, et al. Magnolol inhibits growth of gallbladder cancer cells through the p53 pathway. Cancer Sci 2015; 106: 1341–50.
Bao R, Shu Y, Wu X, et al. Oridonin induces apoptosis and cell cycle arrest of gallbladder cancer cells via the mitochondrial pathway. BMC Cancer 2014; 14: 217.
Li SH, Li CF, Sung MT, et al. Skp2 is an independent prognosticator of gallbladder carcinoma among p27(Kip1)-interacting cell cycle regulators: an immunohistochemical study of 62 cases by tissue microarray. Mod Pathol 2007; 20: 497–507.
Choy B, LaLonde A, Que J, et al. MCM4 and MCM7, potential novel proliferation markers, significantly correlated with Ki-67, Bmi1, and cyclin E expression in esophageal adenocarcinoma, squamous cell carcinoma, and precancerous lesions. Hum Pathol 2016; 57: 126–35.
Kwok HF, Zhang SD, McCrudden CM, et al. Prognostic significance of minichromosome maintenance proteins in breast cancer. Am J Cancer Res 2015; 5: 52–71.
Tomita Y, Imai K, Senju S, et al. A novel tumor-associated antigen, cell division cycle 45-like can induce cytotoxic T-lymphocytes reactive to tumor cells. Cancer Sci 2011; 102: 697–705.
Sun J, Shi R, Zhao S, et al. Cell division cycle 45 promotes papillary thyroid cancer progression via regulating cell cycle. Tumour Biol 2017; 39: 1010428317705342.
Seo YS, Kang YH. The human replicative helicase, the CMG complex, as a target for anti-cancer therapy. Front Mol Biosci 2018; 5: 26.
Sufan RI, Ohh M. Role of the NEDD8 modification of Cul2 in the sequential activation of ECV complex. Neoplasia 2006; 8: 956-63.
Chan CH, Li CF, Yang WL, et al. The Skp2-SCF E3 ligase regulates Akt ubiquitination, glycolysis, herceptin sensitivity, and tumorigenesis. Cell 2012; 149: 1098–111.
Pitroda SP, Wakim BT, Sood RF, et al. STAT1-dependent expression of energy metabolic pathways links tumour growth and radioresistance to the Warburg effect. BMC Med 2009; 7: 68.
Vaughan RA, Garcia-Smith R, Trujillo KA, et al. Tumor necrosis factor alpha increases aerobic glycolysis and reduces oxidative metabolism in prostate epithelial cells. Prostate 2013; 73: 1538–46.
Fields AP, Justilien V. The guanine nucleotide exchange factor (GEF) Ect2 is an oncogene in human cancer. Adv Enzyme Regul 2010; 50: 190–200.
Zhuo YJ, Xi M, Wan YP, et al. Enhanced expression of centromere protein F predicts clinical progression and prognosis in patients with prostate cancer. Int J Mol Med 2015; 35: 966–72.
Takahashi Y, Sheridan P, Niida A, et al. The AURKA/TPX2 axis drives colon tumorigenesis cooperatively with MYC. Ann Oncol 2015; 26: 935–42.
van Gijn SE, Wierenga E, van den Tempel N, et al. TPX2/Aurora kinase A signaling as a potential therapeutic target in genomically unstable cancer cells. Oncogene 2019; 38: 852–67.
Lee CH, Kuo WH, Lin CC, et al. MicroRNA-regulated protein-protein interaction networks and their functions in breast cancer. Int J Mol Sci 2013; 14: 11560–606.
L’Esperance S, Popa I, Bachvarova M, et al. Gene expression profiling of paired ovarian tumors obtained prior to and following adjuvant chemotherapy: molecular signatures of chemoresistant tumors. Int J Oncol 2006; 29: 5–24.
Asteriti IA, Rensen WM, Lindon C, et al. The Aurora-A/TPX2 complex: a novel oncogenic holoenzyme? Biochim Biophys Acta 2010; 1806: 230–9.
Asteriti IA, Daidone F, Colotti G, et al. Identification of small molecule inhibitors of the Aurora-A/TPX2 complex. Oncotarget 2017; 8: 32117–33.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Experimental Oncology
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
