Over-expression of telomere binding factors (TRF1 & TRF2) in renal cell carcinoma and their inhibition by using SiRNA induce apoptosis, reduce cell proliferation and migration invitro

Deeksha Pal; Ujjawal Sharma; Shrawan Kumar Singh; Nandita Kakkar; Rajendra Prasad

doi:10.1371/journal.pone.0115651

Over-expression of telomere binding factors (TRF1 & TRF2) in renal cell carcinoma and their inhibition by using SiRNA induce apoptosis, reduce cell proliferation and migration invitro

PLoS One. 2015 Mar 2;10(3):e0115651. doi: 10.1371/journal.pone.0115651. eCollection 2015.

Authors

Deeksha Pal¹, Ujjawal Sharma¹, Shrawan Kumar Singh², Nandita Kakkar³, Rajendra Prasad¹

Affiliations

¹ Department of Biochemistry, PGIMER, Chandigarh, India.
² Department of Urology, PGIMER, Chandigarh, India.
³ Department of Histopathology, PGIMER, Chandigarh, India.

Abstract

Telomere binding factors viz. TRF1 and TRF2 are a part of sheltrin complex that are present exclusively at the ends of chromosomes. These factors play an important role in maintaining chromosomal integrity at the ends. However, their status and role are not clear in renal cell carcinoma (RCC). Therefore, the present study was conducted to evaluate TRF1 and TRF2 expressions in RCC tissues. Further, the role of these factors involved in tumorigenesis was elucidated by gene silencing using siRNA in RCC cell line (A498). The present study documented a significant over-expression of TRF1 (P = 0.005) and TRF2 (P = 0.0048) mRNAs by real time PCR in RCC tissues as compared with adjacent normal kidney tissues. Immunohistochemistry studies also revealed higher expression of TRF1 and TRF2 proteins in RCC. Moreover, TRF1 or TRF2 gene silencing using siRNA showed marked reduction in proliferation of RCC cells (P = 0.000). Further, significantly induced cell cycle arrest (P = 0.000) and apoptosis of RCC cells (P = 0.000) was documented upon TRF1 or TRF2 gene silencing. Henceforth, the results deduce that TRF1 or TRF2 inhibitions play an important role in the induction of apoptosis in A498 cells, which may serve as a potential therapeutic target in RCC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
Aged
Apoptosis
Carcinoma, Renal Cell / metabolism
Carcinoma, Renal Cell / pathology*
Cell Line, Tumor
Cell Movement
Cell Proliferation
Female
G1 Phase Cell Cycle Checkpoints
Humans
Immunohistochemistry
Kidney / metabolism
Kidney Neoplasms / metabolism
Kidney Neoplasms / pathology*
Male
Middle Aged
RNA, Small Interfering / metabolism*
S Phase Cell Cycle Checkpoints
Telomeric Repeat Binding Protein 1 / antagonists & inhibitors
Telomeric Repeat Binding Protein 1 / genetics
Telomeric Repeat Binding Protein 1 / metabolism*
Telomeric Repeat Binding Protein 2 / antagonists & inhibitors
Telomeric Repeat Binding Protein 2 / genetics
Telomeric Repeat Binding Protein 2 / metabolism*
Up-Regulation

Substances

RNA, Small Interfering
Telomeric Repeat Binding Protein 1
Telomeric Repeat Binding Protein 2

Grants and funding

This is a part of a research project funded by Council of Scientific and Industrial Research, New Delhi, India (vide letter no. 27/0218/09/EMR-II). The authors are thankful to Indian council of medical research, New Delhi, India for awarding junior and senior research fellowship (3/1/3/JRF-2010/HRD-67(11065). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.