| Background and 0bjective Gallbladder cancer is a common malignant tumor of the biliary system,and its incidence rate ranks 6th in digestive tract tumors,accounting for 2/3 of biliary system tumors.Gallbladder cancer is relatively insidious at the onset.There is no clinical symptoms or mild digestive symptoms in the early stage.It is often found during the diagnosis of gallstones or cholecystitis.Once diagnosed,most patients have reached advanced or advanced stage,resulting in distant metastasis.Invasion of the liver,bile duct,pancreatic head,duodenum and transverse colon.Gallbladder cancer has a high degree of malignancy,and the patient's 5-year survival rate is very low.At present,surgical resection is still the only possible cure for gallbladder cancer,but gallbladder cancer surgery is difficult to achieve R0 resection,easy recurrence and metastasis after surgical resection,and gallbladder cancer It is not sensitive to radiotherapy and chemotherapy,and the prognosis is very poor.Therefore,the mechanism of proliferation,invasion and metastasis of gallbladder carcinoma is studied,and effective treatment methods are sought to become the key to prevention and treatment of gallbladder cancer.B7-type molecules are currently a common class of costimulatory molecules,including B7-H3,B7-H1,BTLA,B7-H4,etc.,of which B7-H4(also known as B7S1,B7 X or VTCN1)is a newly discovered negative Co-stimulatory molecules,mainly expressed in malignant tumor cells,found that B7-H4 molecules can inhibit the proliferation and activation of T lymphocytes,reduce the secretion of T lymphocyte factors,regulate the progression of T cell cycle,and lead to the early escape of tumors.The body's immune system is monitored to escape the body's immune attack.Moreover,B7-H4 can promote the proliferation,invasion and metastasis of tumor cells,and plays an important role in the process of tumor development into malignant tumors.Therefore,to study the mechanism of action of B7-H4 molecule in gallbladder cancer,provide more ideas and methods for immunotargeting therapy of gallbladder cancer.In this study,the expression of B7-H4 in human gallbladder carcinoma and gallbladder carcinoma GBC-SD cells was detected by immunohistochemistry.The B7-H4 molecule in gallbladder carcinoma GBC-SD cells was silenced by si RNA and observed for human gallbladder carcinoma GBC-The effect of SD cell growth,proliferation,and invasion ability.Methods T By consulting the medical record system of our hospital,we selected specimens of gallbladder cancer diagnosed by postoperative pathological diagnosis,collected wax specimens,and performed immunohistochemical tests in our pathology department to detect the expression of B7-H4 molecules in gallbladder carcinoma tissues,and selected breasts.The cancer pathological specimens were used as a positive control for immunohistochemistry.The gallbladder carcinoma GBC-SD cells were cultured in parallel and immunohistochemistry was performed to detect the expression of B7-H4 molecules in GBC-SD cells.The human gallbladder carcinoma GBC-SD cells were silenced by small interfering RNA(si RNA).The expression of B7-H4 m RNA was detected by Q-PCR before and after transfection.The expression of B7-H4 protein in GBC-SD cells was detected by Western blot.CCK-8 method was used to observe the effect of GBC-SD cell proliferation on gallbladder carcinoma.Transwell chamber assay was used to detect the invasion ability of gallbladder carcinoma GBC-SD cells.The results were tested for homogeneity of variance and one-way analysis of variance.LSD-was used for comparison.t test,P < 0.05 was considered statistically significant.Results(1)26 cases of pathological sections were immunohistochemically stained,and B7-H4 protein was positively expressed in 18 samples.The positive rate was 69.23%(18/26),and the gallbladder cancer GBC-SD cells B7-H4 The gene translation product was immunohistochemically stained and found that B7-H4 was positively expressed in human gallbladder carcinoma GBC-SD cell specimens,and not expressed or expressed in tissues adjacent to gallbladder carcinoma;(2)B7-H4 si RNA was successfully transfected into human gallbladder carcinoma GBC-SD cells.Q-PCR results showed that B7-H4 m RNA expression level was significantly decreased after si RNA transfection,compared with untransfected group and empty vector group(NC)control group.There was a significant difference(P<0.01);(3)The results of immunoblotting showed that B7-H4 si RNA group significantly inhibited the expression of B7-H4 protein in GBC-SD cells compared with untransfected group and NC group(P<0.01).(4)The proliferation activity of GBC-SD cells transfected with B7-H4 si RNA was lower than that of NC group(P<0.05).(5)Transwell chamber method showed that the number of invasive cells of GBC-SD cells in B7-H4 si RNA group was significantly lower than that in untransfected group and NC group,with significant difference(P<0.01).Conclusions B7-H4 is positively over expressed in human gallbladder carcinoma tissues and gallbladder carcinoma GBC-SD cells,and is low or no expression in gallbladder carcinoma adjacent tissues.B7-H4 si RNA can effectively silence GBC-SD cells and make B7-in gallbladder cancer cells.The expression of H4 molecule m RNA and its protein is decreased,which limits the growth,proliferation and invasion of GBC-SD cells.Therefore,B7-H4 may be a candidate for gene therapy for gallbladder cancer. |
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