| Long-term unresolved gastric cancer,the main reason is the existence of gastric cancer stem cells,and the characteristics of stem cells in different gastric cancer stages are different,and the treatment methods are also not the same.Therefore,in this experiment,we want to explore the difference of stem cell microenvironment in different gastric cancer stages and choose two cell lines: AGS and SGC7901.SGC7901 is derived from gastric adenocarcinoma and is extracted from a patient's lymph node metastases.The AGS cell line also originated from gastric adenocarcinoma but originated from an untreated tumor mass.Since the two cell lines have different migratory and stem cell contents,they can be considered as two different stages: primary gastric cancer and metastatic gastric cancer.The experimental results can distinguish the cells under both metastatic and non-metastatic conditions.The differences in microenvironment can provide clinical guidance for staging of gastric cancer through the comparison of two microenvironments.The topic is to simulate the micro-environment of the hydrogel obtained by cross-linking sodium alginate and hyaluronic acid with calcium chloride and chitosan.Experiments are selected from three aspects: mechanical properties,serum content,and the content and molecular weight of hyaluronic acid.Two gastric cancer cell lines were select by creating a microenvironment that is most suitable for the growth of gastric cancer stem cells in both cell lines and verifying the resulting cell mass.The suitable hydrogel condition for cell line SGC7901 is SA2.2% plus LMW-HA 2.0%;for cell line AGS hydrogel condition is SA 0.6% plus LMW-HA2.0%.Tests revealed that the two cell lines CD44,MDR1,and LGR5 were significantly elevated,among which CD44 was one of the markers of malignancy.Elevation of MDR1 indicates that the resistance of both cell lines was improved,while the high display of LGR5 screened that these cell lines all have gastric stem cell properties.The other markers of SGC7901,NANOG,CD24,and CD133,were significantly elevated,indicating that SGC7901 screened for enhanced cell dryness.The expression of the stem gene expression OCT-4 in the AGS cell line increasedsignificantly,but there was no significant difference in the remaining expression NANOG,CD24,and CD133,indicating that the AGS cell was less dry than the SGC7901.Studies have shown that OCT-4 has a higher expression level in primitive cells,but has a lower level of mature differentiation cells and a lower level of AGS differentiation,resulting in a higher expression level of OCT-4,whereas SGC7901 is a moderately differentiated cell line and therefore OCT-4 Low expression.After the co-culture,the direction of THP1 differentiation was further confirmed as stem cells.After the detection of M1 and M2 markers,the obtained stem cell group can promote the differentiation of THP1 into M2,and further confirm that they are stem cells.This experiment compares the environmental differences of stem cells in different stages of gastric cancer;and the results of co-culture experiments show that gastric cancer stem cells do affect macrophages,and macrophages are an important line of defense for the human immune system.Combining the two can be a new development.The stomach cancer treatment methods and therapeutic drugs provide new ideas. |
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