Anti-tumor and immunomodulatory effects of Hedyotis diffusa Willd.water extract on human breast cancer MDA-MB-231 transplanted tumor in nude mice
Objective It is to investigate the anti-tumor and immunomodulatory effects of Hedyotis diffusa Willd.water extract(HDWE)on human breast cancer MDA-MB-231 xenograft tumor in nude mice,and to explore its mechanism.Methods Six normal BALB/c-nu female nude mice were selected as normal group,another 30 BALB/c-nu female nude mice were used to prepare models of MDA-MB-231 breast cancer transplanted tumor.The successfully modeled mice were randomly divided into model group,cyclophosphamide(CTX)group and HDWE low-dose group and HDWE high-dose group,with 6 mice in each group.The CTX group was given CTX 25 mg/kg by intraperitoneal injection every 7 days,the low and high dose groups of HDWE were gavaged with HDWE 1 500 mg/(kg·d)and 6 000 mg/(kg·d)respectively,and the normal and model groups were gavaged with equal volume of sterilized injectable water,all once daily,continuously treated for 21 days.The activity status of the nude mice was monitored,and their tumor sizes were measured once every 2 days interval.At the end of the treatment,the body weight,tumor weight and spleen weight were measured,the spleen in-dex and tumor suppression rate were calculated;the pathological morphology of the transplanted tumors was observed by HE staining;the proliferative capacity of T and B lymphocytes in spleen was evaluated by splenic lymphocyte proliferation as-say;the number of leukocytes and percentage of lymphocytes in peripheral blood were detected by hematology analyzer;the serum levels of IL-2,IL-6,IFN-γ and TNF-α were detected by ELISA;the expressions of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)pathway proteins and apoptosis-related proteins(Bcl-2,Bax,Caspase-7 and Caspase-9)in tumor tissues of nude mice of the model group and low and high dose groups of HDWE were detected by Western blot assay.Results Compared with the normal group,the spleen index and number of leukocytes in peripheral blood were significantly increased,while the proliferative capacity of T and B lymphocytes in spleen,percentage of lymphocytes in peripheral blood,serum levels of IL-2,IL-6,IFN-γ and TNF-α were significantly de-creased in the nude mice of the model group(all P<0.05),the tumor cells in tumor tissue were densely abundantly ar-ranged in disorder,and their heterogeneity was obvious.Compared with the model group,the spleen index,proliferative capacity of T and B lymphocytes in spleen,percentage of lymphocytes in peripheral blood,and serum levels of IL-2,IL-6,IFN-γ,and TNF-α were significantly increased,while the tumor weight and number of leukocytes in peripheral blood were all significantly reduced in the nude mice of the the low and high dose groups of HDWE(all P<0.05);the number of tumor cells in tumor tissue was greatly reduced,with apoptosis and structural damage;the relative expressions of p-PI3K,PI3K,p-Akt,Akt,mTOR,and Bcl-2 proteins in tumor tissues were significantly reduced(all P<0.05),and the relative expressions of Bax,Caspase-7,and Caspase-9 proteins were significantly increased(all P<0.05).The spleen index,tumor weight,the proliferative capacity of T and B lymphocytes in spleen,the number of leukocytes and percentage of lym-phocytes in peripheral blood in the nude mice of the CTX group were significantly lower than those in the model group(all P<0.05),and the number of tumor cells in tumor tissue was significantly reduced.Conclusion HDWE can significantly inhibit the growth of transplanted tumors,induce apoptosis of breast cancer cells,and enhance immune function of nude mice with MDA-MB-231 breast cancer,the mechanism of which may be related to inhibit the activation of PI3K/Akt/mTOR pathway,down-regulate the expressions of apoptosis-associated proteins,and up-regulate the expressions of pro-ap-optotic proteins.
Hedyotis diffusa Willd.breast cancerMDA-MB-231transplanted tumor in nude miceimmunomodu-latoryapoptosis
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