Expression Of TGF-?1 And TG2 In Cisplatin-treated MCF-7 Breast Cancer Cells And Interrelation

Background:Breast cancer is a metastatic cancer that usually metastasizes to distant organs such as bones,liver,lungs and brain.It is the world's most threatened cancer for women.More than 1.5 million women was diagnosed as breast cancer every year in worldwide.Invasion and metastasis of breast cancer is a very complex process,which involves multiple genes and multiple steps.Therefore,to explore the pathogenesis of breast cancer is the greatest significance for the prevention,treatment and prognosis of breast cancer.TGF-?1 is a secreted cytokine involved in controlling gene expression and ultimately controlling cell cycle and tissue repair.With the occurrence and development of tumor,TGF-?1 shifts from inhibiting cancer cells to promoting cancer cell proliferation,invasion and metastasis.Tissue transglutaminase type 2(TG2)is a multifunctional enzyme that acts on the deamination and acyl transfer reactions of proteins and plays an important role in G protein signaling.Studies have found that TGF-?1 and TG2 are mainly expressed in the matrix and nucleus of cells,and can be transformed in the nucleus and cytoplasm to generate information and play a specific role in transmitting signals.At the same time,it can also play an important role in the metastasis of cancer cells,cell adhesion,and in the process of cell proliferation.However,the role and relationship of TGF-?1 and TG2 in breast cancer has rarely been reported.Cisplatin(DDP)is one of the commonly used chemotherapy drugs for the treatment of gynecological tumors.It belongs to the inorganic metal complex.The ionized chloride ion of cisplatin can cross-link with the DNA in the cancer cells,causing the DNA replication to be blocked,thereby inhibiting the proliferation of tumor cells.In the pathogenesis of breast cancer,whether cisplatin can regulate the expression of TGF-?1 and TG2 has become another focus of this experimental study.Objective:In this experiment,the expression of TGF-?1 and TG2 was explored in breast cancer tissues,and elucidated the role of TGF-?1 and TG2 in breast cancer.The expression of TGF-?1 and TG2 in breast cancer was analyzed by Cisplatin-treated MCF-7 breast cancer cells,and elucidated the targeting effect of Cisplatin on treating breast cancer.After treatment of MCF-7 cells with TGF-?1 Inhibitor and Inducer and TG2 Inhibitor,the expressions of TGF-?1 and TG2 were detected,respectively,and confirmed the relationship between TGF-?1 and TG2.Methods:1.To diagnose the patient's condition by HE staining;to analyze the expression of TGF-?1 and TG2 in breast cancer by immunohistochemical staining.2.The effect of Cisplatin on the morphology of MCF-7 breast cancer cells was observed by microscope.3.The effect of Cisplatin on MCF-7 breast cancer cells was analyzed by CCK-8 assay.4.The effect of Cisplatin on the cell cycle of MCF-7 cells was analyzed by flow cytometry.5.Cellular immunofluorescence was used to analyze the localization and expression of protein in cells.Western blot was used to verify the expression of TG2 and TGF-?1 protein in Cisplatin-treated MCF-7 cells.6.The expression of TG2 after TGF-?1 Inducer induced-TGF-?1 was analyzed by immunofluorescence and Western blot in MCF-7 breast cancer cells.7.Immunofluorescence and Western blot were used to analyze the expression of TG2 protein after TGF-?1 inhibitor-inhibited TGF-?1 in MCF-7 breast cancer cells.Results:1.HE staining results showed that 30 patients were diagnosed with breast cancer;TG2 and TGF-?1 were highly expressed in breast cancer.2.Cisplatin may induce apoptosis of MCF-7 breast cancer cells as compared with untreated breast cancer cells.3.Cisplatin could regulate the cycle in MCF-7 breast cancer cells as compared with untreated breast cancer cells,especially G0/G1.4.The expression of TG2 and TGF-?1 were decreased after Cisplatin-treated MCF-7 cells with untreated breast cancer cells(P<0.05).5.The expression of TG2 and TGF-?1 increased by TGF-?1 inductor-treated-MCF-7 breast cancer cells with untreated breast cancer cells.6.The expression of TG2 might be regulated by TGF-?1 inhibitors-treatedMCF-7 breast cancer cells compared with untreated cell group.Conclusions:1.TG2 and TGF-?1 played an important role in breast cancer.2.Cisplatin could regulate the apoptosis of MCF-7 breast cancer cells and affect the cell cycle of MCF-7 breast cancer.3.The expression of TG2 and TGF-?1 were inhibited by Cisplatin-treated MCF-7 breast cancer cells.4.The expression of TGF-?1 was regulated by Cisplatin or TGF-?1 inhibitor,which might regulate the expression of TG2,and then played a targeted therapeutic role.