Frequent Downregulation and Promoter Hypermethylation of DLC1: Relationship with Clinical Outcome in Gallbladder Cancer

Overview

Journal J Gastrointest Cancer

Publisher Springer

Specialties Gastroenterology
Oncology

Date 2021 Jan 8

PMID 33417200

Authors

Deepika Singh

Amisha Bharti

Dipanjan Biswas

Mallika Tewari

Amrita Ghosh Kar

Mumtaz Ahmed Ansari

Sunita Singh

Gopeshwar Narayan

Affiliations

Soon will be listed here.

Abstract

Introduction: Down regulation of DLC1 is associated with poor prognosis in many cancers, however, its role in gallbladder cancer (GBC) is still unclear. In present study, we investigated the expression profile and promoter methylation status of DLC1.

Methods: Expression profiles of DLC1 in 55 GBC and their paired adjacent control samples were analyzed through real time RT-PCR and immunohistochemistry. The mRNA data was correlated with clinico-pathological parameters. Promoter hypermethylation was analyzed through MSP.

Results: DLC1 shows downregulation in 76.4%, upregulation in 10.9% whereas no change in 12.7% of GBC samples. Its underexpression shows significant correlation with tumor grade and nodal spread. IHC shows cytoplasmic expression of DLC1 in normal as well as tumor samples. IHC result was concordant to mRNA result. Samples having downregulated DLC1 expression show heterozygous methylation in 83.3% of samples and homozygous methylation in 9.5% of samples whereas 7% of samples have no methylation. Kaplan-Meier analysis shows patient with decreased mRNA of DLC1 have significant low mean survival compared to patients with higher mRNA expression of DLC1.

Conclusion: Our findings suggested that dysregulated expression of DLC1 and its hypermethylation may be one of the events playing roles in tumorigenesis of GBC and may serve as a potential target for development of future GBC gene therapy.

References

Sharma A, Sharma K, Gupta A, Yadav A, Kumar A . Gallbladder cancer epidemiology, pathogenesis and molecular genetics: Recent update. World J Gastroenterol. 2017; 23(22):3978-3998. PMC: 5473118. DOI: 10.3748/wjg.v23.i22.3978. View

Wistuba I, Maitra A, Carrasco R, Tang M, Troncoso P, Minna J . High resolution chromosome 3p, 8p, 9q and 22q allelotyping analysis in the pathogenesis of gallbladder carcinoma. Br J Cancer. 2002; 87(4):432-40. PMC: 2376134. DOI: 10.1038/sj.bjc.6600490. View

Wistuba I, Tang M, Maitra A, Alvarez H, Troncoso P, Pimentel F . Genome-wide allelotyping analysis reveals multiple sites of allelic loss in gallbladder carcinoma. Cancer Res. 2001; 61(9):3795-800. View

Liao Y, Lo S . Deleted in liver cancer-1 (DLC-1): a tumor suppressor not just for liver. Int J Biochem Cell Biol. 2007; 40(5):843-7. PMC: 2323245. DOI: 10.1016/j.biocel.2007.04.008. View

Durkin M, Yuan B, Thorgeirsson S, Popescu N . Gene structure, tissue expression, and linkage mapping of the mouse DLC-1 gene (Arhgap7). Gene. 2002; 288(1-2):119-27. DOI: 10.1016/s0378-1119(02)00462-6. View

Yuan B, Miller M, Keck C, Zimonjic D, Thorgeirsson S, Popescu N . Cloning, characterization, and chromosomal localization of a gene frequently deleted in human liver cancer (DLC-1) homologous to rat RhoGAP. Cancer Res. 1998; 58(10):2196-9. View

Clark E, King W, Brugge J, Symons M, Hynes R . Integrin-mediated signals regulated by members of the rho family of GTPases. J Cell Biol. 1998; 142(2):573-86. PMC: 2133065. DOI: 10.1083/jcb.142.2.573. View

Sekimata M, Kabuyama Y, Emori Y, Homma Y . Morphological changes and detachment of adherent cells induced by p122, a GTPase-activating protein for Rho. J Biol Chem. 1999; 274(25):17757-62. DOI: 10.1074/jbc.274.25.17757. View

Jaffe A, Hall A . Rho GTPases in transformation and metastasis. Adv Cancer Res. 2002; 84:57-80. DOI: 10.1016/s0065-230x(02)84003-9. View

10.

Lozano J, Xing R, Cai Z, Jensen H, Trempus C, Mark W . Deficiency of kinase suppressor of Ras1 prevents oncogenic ras signaling in mice. Cancer Res. 2003; 63(14):4232-8. View

11.

Qian X, Durkin M, Wang D, Tripathi B, Olson L, Yang X . Inactivation of the Dlc1 gene cooperates with downregulation of p15INK4b and p16Ink4a, leading to neoplastic transformation and poor prognosis in human cancer. Cancer Res. 2012; 72(22):5900-11. PMC: 7477393. DOI: 10.1158/0008-5472.CAN-12-2368. View

12.

Kim T, Healy K, Der C, Sciaky N, Bang Y, Juliano R . Effects of structure of Rho GTPase-activating protein DLC-1 on cell morphology and migration. J Biol Chem. 2008; 283(47):32762-70. PMC: 2583296. DOI: 10.1074/jbc.M800617200. View

13.

Ullmannova-Benson V, Guan M, Zhou X, Tripathi V, Yang X, Zimonjic D . DLC1 tumor suppressor gene inhibits migration and invasion of multiple myeloma cells through RhoA GTPase pathway. Leukemia. 2008; 23(2):383-90. PMC: 2790147. DOI: 10.1038/leu.2008.285. View

14.

Balmain A, Gray J, Ponder B . The genetics and genomics of cancer. Nat Genet. 2003; 33 Suppl:238-44. DOI: 10.1038/ng1107. View

15.

Wang D, Qian X, Rajaram M, Durkin M, Lowy D . DLC1 is the principal biologically-relevant down-regulated DLC family member in several cancers. Oncotarget. 2016; 7(29):45144-45157. PMC: 5216712. DOI: 10.18632/oncotarget.9266. View

16.

Jaffe A, Hall A . Rho GTPases: biochemistry and biology. Annu Rev Cell Dev Biol. 2005; 21:247-69. DOI: 10.1146/annurev.cellbio.21.020604.150721. View

17.

Braun A, Olayioye M . Rho regulation: DLC proteins in space and time. Cell Signal. 2015; 27(8):1643-51. DOI: 10.1016/j.cellsig.2015.04.003. View

18.

Yuan B, Zhou X, Durkin M, Zimonjic D, Gumundsdottir K, Eyfjord J . DLC-1 gene inhibits human breast cancer cell growth and in vivo tumorigenicity. Oncogene. 2003; 22(3):445-50. DOI: 10.1038/sj.onc.1206064. View

19.

Yuan B, Jefferson A, Baldwin K, Thorgeirsson S, Popescu N, Reynolds S . DLC-1 operates as a tumor suppressor gene in human non-small cell lung carcinomas. Oncogene. 2003; 23(7):1405-11. DOI: 10.1038/sj.onc.1207291. View

20.

Guan M, Zhou X, Soulitzis N, Spandidos D, Popescu N . Aberrant methylation and deacetylation of deleted in liver cancer-1 gene in prostate cancer: potential clinical applications. Clin Cancer Res. 2006; 12(5):1412-9. DOI: 10.1158/1078-0432.CCR-05-1906. View