The Prevention And Treatment Efficacy Of Coix Seed Oil On Lung Cancer-promoting Effect Of Obesity Microenvironment And Its Mechanisms

Lung cancer is the leading cause of cancer death in both more and less developed countries.Due to serious air pollution recently in China,there has been the highest lung cancer incidence in the world.Undoublely,tumor prevention is the best way for reducing the burden of lung cancer screening,treatment,and mortality,whereas knowing lung cancer-related risk factors is a precondition for successful tumor prevention.A lot of evidences showed that obesity is a risk factor for cancer and cancer-related mortality,however,its role in lung cancer progression remains controversial.Coix seed oil is an active ingredients extracted from traditional Chinese medicine semen coicis and its commercial agent is Kanglaite Injection which is mainly used for inoperable primary non-small cell lung cancer and primary hepatic carcinoma.Kanglaite Injection combined with chemotherapy can improve response rate,life quality,symptoms,and reduce incidence of side effects.Other reports showed that Coix seed oil can reduce the abdominal fat tissue.Considering tumor-promoting effect of obesity microenvironment-related inflammation,we guess that the fat-reducing effect of Coix seed oil should prevent the lung cancer-promoting effect of obesity microenvironment if.obesity can promote lung cancer progression.The aim of this study is to confirm whether obesity microenvironment promotes lung cancer progression and whether this promotion can be decreased by Coix seed oil.This subject includes in vitro study,in vivo study and action mechanism.In vitro,after treatment with different concentrations of Coix seed oil on cells,MTT assay was used to test the proliferation ability of 3T3-L1,HUVEC and LLC cells;cell scratch assay was carried to test the migration ability of LLC cells;cell permeability was detected by evens blue infiltration in HUVEC cells;AO/EB staining and oil red O staining were used to detect the apoptosis and differentiation of 3T3-L1 cells,respectively.In vivo,three lung cancer models(urethane-induced lung adenocarcinoma model,LLC allograft model and orthotopic lung cancer model)were established in C57BL/6J and KKay mice.Mice were fed standard rodent chow(SRC)or high-fat diet(HFD)and received Coix seed oil emulsion by lavage everyday.The physiological index include body weight,autonomic activities,carinogenic incidence and tumer multiplicity of mice were observed.The pathologic changes in mice were assessed by HE staining.Insulin and glucose tolerance test were performed using an ELISA assay kit in urethane-induced lung adenocarcinoma model mice.Lung epithelial proliferation and epithelial-mesenchymal transition(EMT)of tumor tissue were examined by immunohistochemical analysis in urethane-induced lung adenocarcinoma model and LLC orthotopic lung cancer model to evaluate whether overweight or obesity contributes to carcinogenesis.To further explore the mechanism of lung cancer prevention and cure effect of Coix seed oil,the serum concentrations of insulin,adiponectin,leptin,cholesterol,Interleukin-6(IL-6),Super C-reactive protein(hs-CRP),tumer necrosis factor(TNF-?)as well as Reactive Oxygen Species(ROS)and 8-hydroxy-deoxyguanine(8-OHd G),markers of oxidative stress,and hydroxy indole acetic acid(5-HIAA),a marker of vascular disruption,were determined using an ELISA assay kit according to the manufacturer's protocols.The intergrity of the lung epithelium was assayed using the Evens blue extra-barrier technique and lung water content was measured by dry-wet method in a urethane-induced lung adenocarcinoma model.Overweight-or obesity-associated changes in cellular signaling molecules,including janus kinase(JAK),signal transducers and activator of transcription 3(STAT3),protein kinase B(Akt),mammalian target of rapamycin(m TOR),nuclear factor-?B(NF-?B)and Cyclin D1 were examined by Western blot analyses in a urethane-induced lung adenocarcinoma model.In vitro study shows,Coix seed oil significantly suppressed the proliferation of 3T3-L1 preadipocyte cells but seemed no obvious inhibition effect on the proliferation of HUVECs and LLCs.Coix seed oil had no significant changes on migration of LLCs,however,it attenuated endothelial cell permeability of HUVECs and induced apoptosis and differentiation of 3T3-L1 s.In vivo,the experimental results shows,compared to the same strain of mice fed the SRC,HFD-fed C57BL/6J mice was overweight and HFD-fed KKay mice was obesity.Two strains of HFD diet mice showed excessive weight gain and a decrease in autonomic activity,moreover,the increased carcinogenic incidence and tumer multiplicity in urethane-induced lung cancer model and orthotopic lung cancer model mice as well as increased tumer weight and lung metastatic nodes in LLC allograft model.HFD diet also caused an abnormal in lung pathology.The results above lead to a shorten survival period and higher mortality in HFD-fed mice.However,after long-term treatment of Coix seed oil,two strains of HFD-fed mice showed weight loss,an increase in autonomic activity,decreased carcinogenic incidence and tumer multiplicity in urethane-induced lung cancer model and orthotopic lung cancer model mice as well as decreased tumer weight and lung metastatic nodes in LLC allograft model.It also improved the abnormal of lung pathology.Mechanism research shows,the insulin and glucose tolerances,serum concentrations of insulin,adiponectin,leptin,cholesterol,IL-6,hs-CRP,TNF-?,as well as ROS,8-OHd G and 5-HIAA in the two HFD-fed strains of mice were incresaed.Besides,lung epithelial integrity and lung water content was increased,the expression of incellular signaling molecules include JAK,STAT3,Akt,m TOR,NF-?B and cyclin D1 was increased in the two strains of HFD-fed mice.These changes promoted the the proliferation of lung epithelial cells and EMT in the process of carcinogenesis in urethane-induced lung cancer model mice.After long-term treatment of Coix seed oil,the obesity-related energy imbalance and metabolic dysfunction was improved,the physiological changes of tissue and cells which are beneficial to the development of tumor are inhibited,the proliferation of lung epithelial cells and the epithelial mesenchymal transition were prevented in the process of carcinogenesis in the two strains of HFD-fed mice.That's all lead to an extended survival and a reduced mortality in mice,eventually.In conclusion,obesity microenvironment plays an promotion role on the development of lung cancer.However,the treatment of Coix seed oil can significantly improve the obesity microenvironment-induced cancer-promoting effect.The mechanism of action is to improve the energy imbalance,metabolic disorder and the increase of interstitial fluid.