Study On CA215 And Its Monoclonal Antibody RP215 In Colorectal

[Background and Objectives]CA215 is a glycoprotein-related antigenic epitope in a glycoprotein, and CA215 is homologous to the heavy chain of human immunoglobulin. In the normal human immunoglobulin and human tissue almost no expression, but in most of the tumor tissue expression. The present literature suggests that CA125 and its monoclonal antibody RP215 can act as immunodiagnostic markers for a variety of tumors and develop immunotherapeutic drugs targeting antibodies.The purpose of this study was to investigate the expression of CA215 in colorectal cancer tissues and serum. To analyze the relationship between CA215 and colorectal cancer invasion, metastasis and prognosis, to evaluate whether CA215 could be used as a suitable tumor Markers for the diagnosis and efficacy of colorectal cancer. To study the effect of RP215 on immunotherapy of colorectal cancer cell lines.Methods:1. Immunohistochemical method was used to detect the expression of CA215 in 132 cases of colorectal cancer. The relationship between the expression of CA215 and the clinicopathological data was analyzed by CA215 positive staining and negative staining.2. The level of serum CA215 in serum of 83 healthy people, 15 patients with IBD, 20 patients with colonic polyps and 103 patients with colorectal cancer was determined by ELISA (serum CA215 level> 11.35 AU / ml was defined as positive) Feasibility of serum markers in colorectal cancer patients. To compare the specificity and sensitivity of CEA and CA199 in colorectal cancer with colorectal cancer.3. The expression of CA215 in colorectal cancer cell lines SW480, Caco2, HCT116 and Lovo and normal intestinal epithelial cells in NCM460 was detected by Western blotting. CA215 was detected by immunofluorescence in NCM460, SW480 and HCT116 The expression of the situation.4. The effects of RP215 on proliferation and apoptosis of colorectal cancer cell line SW480, HCT116 and human normal colonic epithelial cell line NCM460 were measured by CCK8 cell proliferation assay and flow cytometry respectively.Results:1. 132 patients with colorectal adenocarcinoma,CA215 positive (immunohisto-chemical score ? 1) in patients with 68 cases, the positive rate was 51.5%.CA215 negative (immunohistochemical score <1 points) in a total of 64 patients, accounting for 48.5%. The expression of CA215 was associated with local infiltration, histological differentiation, lymph node metastasis and distant metastasis (P <0.05) in patients with colorectal cancer. But not with age, sex and tumor size in patients with colorectal cancer (P> 0.05).2. The results of serum ELISA showed that there was no significant difference between the Crohn's group and the colonic polyps group and the healthy control group (p> 0.05). Compared with the healthy control group, there was asignificant difference between the two groups Significant difference (P <0.01). In addition, there were significant differences in the levels of serum CA215 (P<0.01) in patients with colorectal cancer at different stages (stage?, stage ?) compared with those in stage (stage ?, ?) The higher the serum CA215 level. But there was no significant correlation between early stage(stage ? and stage ?) and late stage (stage ? and stage IV) (p> 0.05). At the same time in the colorectal cancer can also be detected significantly increased CA215 level. The detection rate of CA215 in colorectal cancer was nearly 50%, which was higher than that of CEA (38%) and CA199 (16%). At the same time CA215 combined with CEA detection can improve the detection rate of colorectal cancer to 56-94%.3. Western Blot showed that CA215 was low in normal colon epithelial cells NCM460 and positive expression in SW480, Caco2, HCT116 and Lovo, and the expression of SW480 was the highest The results of immunocyto-chemistry showed that CA215 was low in normal colonic epithelial cells(NCM460) and was expressed in SW480 and HCT116. The expression of CA215 was mainly expressed in cell membrane and cytoplasm.4. The results of CCK8 cell proliferation assay showed that RP215 could inhibit the proliferation of colorectal cancer cell lines SW480 and HCT116 in three concentrations (100ng / ml, 1?g / ml, 10?g / ml), and had a dose-dependent and time-dependent The higher the concentration, the longer the action time,the stronger the inhibitory effect on the proliferation of colorectal cancer cells.And for the normal intestinal epithelial cell line NCM460, 3 concentrations of RP215 can not inhibit its value. The results of apoptosis detection showed that RP215 could induce the apoptosis of colorectal cancer cell lines SW480 and HCT116 at 3 concentrations (0.1 ?g / ml, 1?g/ ml, 10?g / ml). The higher the concentration of RP215, The stronger the ability of apoptosis. But not statistically significant (P> 0.05). In the normal intestinal epithelial cell line NCM460 RP215 can not induce apoptosis.Conclusions:1. The expression of CA215 in patients with colorectal cancer was correlated with local infiltration, histological differentiation of tumor, lymph node metastasis and distant metastasis. At the same time, serum CA215 level and tumor stage (early and late stage ), Suggesting that CA215 may serve as a biomarker for the invasion, metastasis and poor prognosis of patients with colorectal cancer.2. Compared with CEA,CA199, CA215 has a higher detection rate,and CA215 combined with CEA can further improve the diagnosis of colorectal cancer patients. CA215 is more suitable for the early diagnosis of colorectal cancer because CA215 is higher in early detection rate than CEA and CA199, and CEA in advanced colorectal cancer is higher than CA215.CA215 monoclonal antibody RP215 can inhibit the proliferation of colorectal cancer cells,promote tumor apoptosis, and based on CA215 monoclonal antibody may be a new drug for colorectal cancer immunotherapy.3. CA215 monoclonal antibody RP215 can inhibit the proliferation of colorectal cancer cells, induce tumor apoptosis, so based on CA215 monoclonal antibody may be a new drug for colorectal cancer immunotherapy.