Transcriptional Regulation of Epithelial to Mesenchymal Transition Related Genes by Lipopolysaccharide in Human Cervical Cancer Cell Line HeLa

Abstract

Objective: Cancer is one of the common diseases in the world, and cervical cancer is the fourth one. In this type of cancer, many risk factors, especially infectious diseases, such as human papilloma virus (HPV) and gram-negative bacteria can have important effects on the expression of epithelial to mesenchymal transition related genes like Snail, E-cadherin, and ZEB-1, responsible for connecting cell tissues. In this study, we have investigated the effect of Escherichia coli O111:B4 Lipopolysaccharide (LPS) on HPV positive cell line (HeLa), the expression level of the (Snail, E-cadherin, and ZEB-1), HPV oncogenes (E6, E7) and also microRNA-9, 192. Materials and Methods: HeLa cell line was treated with LPS to analyze Snail, E-cadherin, ZEB-1, E6, E7 and also microRNA-9, 192 expression by quantitative real-time PCR in 24, 48 and 72 hours. Results: Quantitative real-time PCR revealed a significant reduction in E-cadherin mRNA level at 10ug/L of LPS in three time-points and after 24 hours at 5ug/L of LPS; however, ZEB-1 at 10ug/L of LPS and Snail at 5, 10ug/L of LPS are up-regulated. E7 also illustrated a slight increase, but we did not find any relationship between E7 and LPS treatment. Additionally, there are upward trends in microRNA-9, 192 levels. Conclusion: The result of this study, LPS is able to reduce E-cadherin expression, caused by increase in repressor E-cadherin protein expression and some microRNAs, probably. Since bacterial infection can be in cervical site, it is likely to be effective in reducing the E-cadherin expression in the EMT and enhance cancer process, therefore; removing these infections by using the appropriate antibiotics may result in slowing down this process, which requires more research.

Keywords: E-cadherin; HPV; LPS; Snail; ZEB-1.

Figure 1

HeLa Cell Line was Treated with LPS by 1, 5 and 10 mmol/L, when the Cells Reached 80% Confluency. In all times of 10 mmol/L of LPS and 24 hours after treating with 5 mmol/L of LPS, E-cadherin expression have been raised

Figure 2

HeLa Cell Line have been Cultured with DMEM Containing LPS at Concentration of 1, 5 and 10 mmol/L. Snail Expression was Revealed a Remarkable Growth in 5 and 10 mmol/L of LPS

Figure 3

LPS is Able to Increase Expression of ZEB-1, in HeLa Cell Line, after 48 and 72 hours in 10 mmol/L

Figure 4

The Level of E7 HPV Expression after Treating HeLa Cell Line with LPS .The bar graph have demonstrated a meaningful increase in amount of E7 HPV expression, however it is not meaningful relation

Figure 5

There are a Remarkable Increase in the amount of microRNA-9 and 192 Expressions after Treating LPS to HeLa Cell Line