Effects of Different Molecular Weight Hyaluronic Acid on the Transcriptome of Inflammation-related Colon Cancer Cells

Abstract

In order to study the main signaling pathways and the expression rules of key genes differentially expressed by different molecular weight hyaluronic acid on human colon cancer cells, Chinese hamster ovary cells (CHO) and pMH3 were used in this study to express the chimeric hyaluronidase PH20, and obtain stable expression and expression. Hyaluronic acid fragment HA35 was prepared by enzymatic digestion with high-purity PH20. Human peripheral blood leukocytes and human colon cancer cell line (HT-29) were used to establish inflammation-related colon cancer cell models, and they were randomly divided into control group and experimental group. The control group was cultured with complete medium for 24 h, in the experimental group, 20 μg/mL of high-molecular-weight hyaluronic acid (HA) and complete medium of HA35 were added for 24 h, and the effect of hyaluronic acid on the expression of TNF-α in inflammation-related colon cancer cells was detected by enzyme-linked immunosorbent assay (ELISA). Human colon cancer cells were collected and total RNA was extracted, reverse-transcribed to establish a cDNA library, and high-throughput sequencing was performed using an Illumina Hiseq X Ten sequencer to establish a transcriptome analysis platform at the mRNA level of HT-29 cells.Transcriptome analysis showed that there were 1372 differentially expressed genes in the HA treatment group compared with the control group, and 966 differentially expressed genes in the HA35 treatment group compared with the control group. These differential genes were mainly related to Rap1, Ras, PI3K-Akt, TGF-β and other inflammatory signaling pathways are related, and differential expression analysis found that HA may inhibit the further development of colon cancer cells by reducing the expression of inflammatory factors such as TNF, IL-17, and IL-22. This study can provide a reference for in-depth exploration of the molecular response mechanism of hyaluronic acid to inflammatory changes in colon cancer cells.