Effect Of Platelets On Biological Function Of Hepatocellular Carcinoma HepG2 Cells And SMMC7721 Cells

backgroundHepatocellular carcinoma（HCC）is the most common malignant liver disease in the world,and more than half of the new cases and deaths are in China,which seriously affects the health of Chinese people.More and more evidence has shown that platelets play a key role in the maintenance of liver homeostasis as well as in the pathological and physiological processes of various liver diseases.Platelets not only participate in hemostasis,activated platelets can also produce cytokines and interact with hepatocytes and tumor microenvironment through its responsible signaling pathway to promote the occurrence and development of tumors.Previous clinical studies have shown that platelets are negatively correlated with the prognosis of patients with liver cancer,and positively correlated with the formation of tumor thrombectomy.These studies suggest that platelets may play a significant role in the development of hepatocellular carcinoma.Platelets are abundant cell types in peripheral blood,which can be easily separated and counted.It is of great clinical significance to predict the prognosis of patients with liver cancer by monitoring platelet counts.However,the role of platelets in the development of HCC has not been clarified.Therefore,it is of great significance to study the role of platelets and their components in the occurrence and development of hepatocellular carcinoma and inflammatory tumor microenvironment,which will provide a theoretical basis for finding targets that effectively block platelet-tumor cell interaction without affecting the normal physiological function of platelets.ObjectiveTo study the role of platelets in the occurrence and development of hepatocellular carcinoma（HCC）and inflammatory tumor microenvironment by observing the effects of intact platelets and broken platelets on the proliferation,apoptosis,invasion and expression of inflammatory factors in Hep G2 cells and SMMC-7721 cells.Methods1.The co-culture model of PLT/PL/PRP/PPP and HCC cells was established:EDTA-K₂ anticoagulant blood was collected from normal people,platelets（PLT）were isolated,and platelet-rich plasma（PRP）,platelet-poor plasma（PPP）and platelet lysate（PL）were prepared.And HCC cells Hep G2 cells or SMMC-7721I cells were cultured with DMEM medium containing different numbers of platelets.2.The effect of platelets on the biological function of HCC was detected:CCK-8method was used to detect the effect of platelets on the proliferation of cells Hep G2 cells and SMMC7721 cells.The effect of platelets on apoptosis and invasion ability of hepatocellular carcinoma cells Hep G2 and SMMC7721 was detected by flow cytometry and Transwelltechnology,respectively.And the effect of platelets on the expression of EMT markers such as E-cadherin,Fibranectin,N-cadherin,Snail,Vimentin,ZEB-1 andβ-cateninin Hep G2 and SMMC7721 cells was detected by RT-PCR assay.3.The effect of platelets on the inflammatory tumor microenvironment of HCC was detected:RT-PCR assaywas used to detect the effect of platelets on the secretion of inflammatory cytokines（IL-1β,IL-6,IL-8,TNF-α,i NOS）and chemokines（CCL2,CCL5）deriving from Hep G2 and SMMC7721 cells.Results1.Platelets and their lytic components can promote the proliferation of Hep G2 cells and SMMC7721 cells.2.Platelets and their lytic components can inhibit the apoptosis of Hep G2 cells and SMMC7721 cells.3.Platelets and their lytic components can promote the invasion of Hep G2 cells and SMMC7721 cells.4.The expression of mesenchymal cell markers（Fibranectin,N-cadherin,Snail,Vimentin,ZEB1,β-catentin）in Hep G2 and SMMC7721 cells was affected by Platelets and their lytic components,which significantly promoted the EMT transformation of HCC cells.5.Platelets and their lysate components can affect the expression of inflammatory cytokines（IL-1β,IL-6,IL-8,TNF-α,i NOS）and chemokines（CCL2,CCL5）in Hep G2cells and SMMC7721 cells to varying degrees,and promote the formation of inflammatory microenvironment of HCC.ConclusionPlatelets play an important role in the progression of HCC,which can significantly promote the occurrence and development of HCC and the formation of inflammatory tumor microenvironment.