Helicobacter pylori-Induced Inflammation Microenvironment Promotes Colon Cancer SW620 Cell EMT
SONG Guangyong1, ZHANG Xiaorong2, LI Mengjun2, GAO Yanping2*
This article explores the effects of HP inflammation microenvironment on EMT of colon cancer SW620 cells. The ELISA method was used to detect the changes of inflammatory factors MIF and IL-1β in the supernatant after HP intervention in U937 cells. RT-PCR was used to detect the changes of MIF, IL-1β, NF-κB gene levels. Scratch assay was used to detect the invasion and migration ability of SW620 cells after treatment with in flammatory supernatant. Effect of inflammatory supernatant on proliferation of SW620 cells after HP-treated U937 cells was observed by CCK-8 method. Changes of EMT-related proteins Vimentin, E-cadherin and N-cadherin in SW620 cells treated with LPS supernatant were detected by Western blot assay. Immunofluorescence assay was used to detect the fluorescence changes of EMT-related protein Vimentin in SW620 cells treated with inflammatory supernatant. Western blot was used to detect the changes of NF-κB protein in U937 cells treated with HP, and EMT-related protein Vimentin, E-cadherin and N-cadherin in SW620 cells treated with inflammatory supernatant. The results showed that HP could increase the expression of MIF, IL-1β and NF-κB in U937 cells. Scratch test showed that the invasion and migration ability of SW620 cells increased with the passage of time points at 12 h, 24 h and 36 h. Immunofluorescence test showed that Vimentin green dot aggregation of SW620 cells increased significantly. Western blot analysis showed that the expression of NF-κB protein in U937 cells was significantly increased at 16 h and 24 h. There was no significant change in the expression of Vimentin protein after intervention of SWPS cells in LPS supernatant, the expression of N-cadherin protein in 24 h group was decreased and E-cadherin protein was increased. The expression of Vimentin protein in SW620 cells increased significantly and the expression of E-cadherin and N-cadherin protein decreased significantly at 12 h, 24 h and 36h after intervention of HP inflammatory microenvironment supernatant. The results of this experimental study indicated that H. pylori-induced microenvironment can promote the occurrence of EMT in SW620 cells .
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