Alendronate blocks human cholangiocarcinoma cell proliferation and migration

Benjaporn Buranrat,Auemduan Prawan,Veerapol Kukongviriyapan,Laddawan Senggunprai

doi:10.4314/tjpr.v18i6.5

Abstract

Purpose: To explore the effect of alendronate on cell death and migration of cholangiocarcinoma (CCA). Methods: Migration and cell death of CCA cells were determined using sulforhodamine B (SRB), colony formation, wound healing, and gelatin zymography assays. The mechanism of action of alendronate was studied with reverse-transcriptase polymerase reaction (RT-PCR) for gene expression and by Western blotting analysis for protein expression. Results: Alendronate stimulated KKU-100 cell death in dose- and time-dependent manner, with low IC50 value, and significantly inhbited colony formation at doses of 5 - 100 µM. Moreover, alendronate at doses of 250 - 1000 µM significantly stimulated CCA apoptosis via reactive oxygen species (ROS) generation, and enhanced caspase 3 activity at a dose of 1000 µM. Moreover, at a dose of 250 µM, it significantly inhibited cell growth through induction of caspase 3 and p53, and reduction of protein expression levels of NF-ĸB. Furthemore, alendronate altered mevalonate (MVA) pathway via downregulation of Rac1 protein expression. In contrast, it significantly inhibited CCA cell migration, and reduced MMP 2 and MMP 9 levels at doses of 25 - 100 µM. Conclusion: Alendronate may be useful as a novel drug for prevention and chemotherapy of CCA.

Highlights

Bisphosphonates (BPs) are mevalonate (MVA) pathway inhibitors which generally inhibit farnesyl pyrophosphate synthase (FPPS), thereby inhibiting the prenylation of small signaling proteins for example Rab, Ras, and Rho [1].Many reports have shown that BPs suppress cancer cell growth and stimulate apoptosis in myeloma, breast, pancreas, liver, ovarian, and prostate cancerous cells [2]
Alendronate inhibited KKU-100 cell growth in concentration- and time-dependent manners, relative to the untreated control group (Figure 1 A and 1 B), with IC50 value of 736.10 ± 136.44 μM after 24 h, and 360.19 ± 36.52 μM after 48 h
The cell were exposed to 250 μM of alendronate and subjected to analysis of apoptosis

Summary

Introduction

Bisphosphonates (BPs) are mevalonate (MVA) pathway inhibitors which generally inhibit farnesyl pyrophosphate synthase (FPPS), thereby inhibiting the prenylation of small signaling proteins for example Rab, Ras, and Rho [1]. Many reports have shown that BPs suppress cancer cell growth and stimulate apoptosis in myeloma, breast, pancreas, liver, ovarian, and prostate cancerous cells [2]. Alendronate is a BP which is used for treating osteoporosis as well as cancer. It has been indicated that alendronate and other BPs suppress prostate cancer invasion. [3], decrease human epidermal carcinoma cells proliferation [4], reduce angiogesis [5]. BPs exert their activities on cancer cells by acting on telomerase activity [6], mevalonate (MVA) pathway [7], and by reducing the protein-related proliferation. Not much is known the effect of alendronate on cholangiocarcinoma (CCA)

Results

Discussion

Conclusion