Expression and Functionality of Connexin-Based Channels in Human Liver Cancer Cell Lines.

Bruna Dos Santos Rodrigues,Mathieu Vinken,Andrés Tabernilla,Bruno Cogliati,Cícero Júlio Silva Costa,Axelle Cooreman,Alanah Pieters,Raf Van Campenhout,Kaat Leroy

doi:10.3390/ijms222212187

Abstract

Liver cancer cell lines are frequently used in vitro tools to test candidate anti-cancer agents as well as to elucidate mechanisms of liver carcinogenesis. Among such mechanisms is cellular communication mediated by connexin-based gap junctions. The present study investigated changes in connexin expression and gap junction functionality in liver cancer in vitro. For this purpose, seven human liver cancer cell lines, as well as primary human hepatocytes, were subjected to connexin and gap junction analysis at the transcriptional, translational and activity level. Real-time quantitative reverse transcription polymerase chain reaction analysis showed enhanced expression of connexin43 in the majority of liver cancer cell lines at the expense of connexin32 and connexin26. Some of these changes were paralleled at the protein level, as evidenced by immunoblot analysis and in situ immunocytochemistry. Gap junctional intercellular communication, assessed by the scrape loading/dye transfer assay, was generally low in all liver cancer cell lines. Collectively, these results provide a full scenario of modifications in hepatocyte connexin production and gap junction activity in cultured liver cancer cell lines. The findings may be valuable for the selection of neoplastic hepatocytes for future mechanistic investigation and testing of anti-cancer drugs that target connexins and their channels.

Highlights

Liver cancer, mainly represented by hepatocellular carcinoma (HCC), is the second and sixth most common cause of cancer death among men and women, respectively [1].Primary liver cancer is associated with a wide variety of causes, including chemical, viral and dietary factors [2,3]
Liver cancer cell lines are frequently used in vitro tools to test candidate anti-cancer agents as well as to elucidate mechanisms of liver carcinogenesis [25], such as cellular communication mediated by connexin-based gap junctions [13]
Many cell lines have a carcinogenic background and are frequently relied upon to test candidate anti-cancer drugs, as well as to elucidate mechanisms of carcinogenesis [51]. Among such mechanisms is cellular communication mediated by gap junctions [13,52]

Summary

Introduction

Mainly represented by hepatocellular carcinoma (HCC), is the second and sixth most common cause of cancer death among men and women, respectively [1].Primary liver cancer is associated with a wide variety of causes, including chemical, viral and dietary factors [2,3]. Mainly represented by hepatocellular carcinoma (HCC), is the second and sixth most common cause of cancer death among men and women, respectively [1]. While some animal models of liver cancer can provide a good reflection of the corresponding human disease and have high translational value, they come with a number of issues [7,8,9,10]. In this respect, besides the obvious ethical constraints, animal models are not always well-fit for mechanistic investigation because of their complex dynamic nature [11]

Methods

Results

Conclusion