| Objective: To investigate the clinical significance of CCL5/CCR5 in gastric cancer by collecting the clinical data and detecting the expression of CCL5/CCR5 in gastric cancer and adjacent tissues,and to explore the role of chemokine CCL5 and its receptor CCR5 in the proliferation,invasion and migration of gastric cancer cell SGC7901 and BGC823,so as to provide more basis for the prevention,diagnosis and treatment of gastric cancer.Method: Part 1: 55 cases patients underwented laparoscopic radical gastrectomy with completed clinical and pathological files of cancer and adjacent tissue were detected by immunohistochemical method(SP).The immunohistochemical staining results were determined by the histologic Scoring System with two pathologists.The expression of CCL5/CCLR5 and its clinicopathological signifincance were analyzed by ?2 test or consistency check.Part 2: Gastric cancer cell lines SGC-7901 and BGC-823 were selected for Westernblot to detect the expressions of CCL5 and CCR5 in two gastric cancer cell lines,so as to determine the influence of CCL5 and CCR5 in the experiment.The cell counting kit-8(CCK-8)test detected the effects of different drug concentrations of CCL5 on the proliferation capacity of SGC-7901 and BGC-823,in order to obtained the appropriate concentration,and to detect the effect of different concentrations of Maraviroc(CCR5 antagonist)on the proliferation ability of gastric cancer cells cultured with quantitative concentration of CCL5.Then cell scratch assay and Transwell invasion assay were used to detect the influence of CCL5 and Maraviroc on the migration ability and invasion ability of SGC-7901 and BGC-823 cells.Result:1.CCL5 was positively expressed in gastric cancer tissues,with a positive expression rate of 90.9%.The expression level was closely related to the differentiation degree,infiltration depth,TNM stage,lymph node metastasis and intravitary tumor thrombus of gastric cancer(P<0.05),while other clinical andpathological characteristics were not significantly correlated(P>0.05).CCR5 was positively expressed in gastric cancer tissues,with a positive expression rate of85.45%.The expression level was correlated with differentiation degree,tumor size,infiltration depth,TNM stage,lymph node metastasis and intravascular tumor thrombus of gastric cancer(P<0.05),but not correlated to other clinical and pathological characteristics(P>0.05).Spearman rank correlation analysis showed that there was a strong correlation between the expression of CL5 and CCR5(rs=0.652,P<0.001),and the expression was correlated with the prognosis of the patients(P<0.05).2.The expression of CCL5 in the gastric cancer cell lines SGC-7901 and BGC-823 detected by Western blot was small,and there was not difference statistically significant between the expression of two cells(P<0.05).Considering that the secretion of CCL5 by gastric cancer cells themselves had little influence on the experiment.It was believed that the CCL5 secretion of cancer cells themselves had little influence in the experiment.The expression level of CCR5 in SGC-7902 and BGC-823 was high,and the comparison between the two cells showed that the expression level of CCR5 in BGC-823 cell line was higher,the difference was statistically significant(P<0.05).3.When CCL5 was used to culture gastric cancer cell line SGC-7901,the drug concentration was above 25ng/ml,and the proliferation ability was significantly enhanced(P<0.05).The proliferation ability was further increased with the increase of drug concentration(24h F=60.131;48 h F = 20.436;72 h F = 24.415).Similarly,when the gastric cancer cell line BGC-823 was cultured,the drug concentration was more than 12.5ng/ml,and the proliferation ability was significantly enhanced(P<0.05).The proliferation ability increased with the increase of drug concentration(24h F=40.214).48 h F= 31.482;72 h F= 45.9411).When 50ng/ml CCL5 gastric cancer cell line SGC-7901 was cultured in Maraviroc with concentration gradient,the drug concentration was above 200ug/ml,and the proliferation ability was significantly inhibited(P<0.05).The inhibitory ability was further increased with the increase of drug concentration(24h F=60.131).48 h F = 20.436;72 h F = 24.415).Similarly,when the cell line BGC-823 was cultured,the drug concentration was above 100ug/ml,and the proliferation ability was significantly inhibited(P<0.05).The inhibitory ability increased with the increase of drug concentration(24h F=21.521;48 h F = 105.207;72 h F= 442.108).4.When SGC-7901 was cultured in extracellular CCL5(50,100ng/ml),the cell migration and invasion abilities were significantly enhanced(P<0.05).As well as gastric cancer cells lines SGC-7901 and BGC-823 were cultured with Maraviroc(200,400ug/ml),the migration ability and invasion ability of gastric cancer cells were decreased(P<0.05).Conclusion:1.The expression level of CCL5 and CCR5 in gastric cancer tissues is closely related to the degree of differentiation,depth of infiltration,TNM stage,lymph node metastasis,intravascular tumor thrombus,and prognosis.2.In vitro experiments,CCL5 and CCR5 promote the proliferation,infiltration and migration of gastric cancer cells,and antagonistic CCR5 may be a new target for the treatment of gastric cancer. |
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