| Lung cancer is the world’s most common malignant tumor which occurs in bronchial epithelium.Its incidence and mortality rate increased year by year.Besides,the cause of lung cancer is complicated.Due to our country industrialization and urbanization fast development in recent years,the air pollution is serious in some places,international agency for research on cancer found that higher PM2.5 in the environment were positively correlated with lung cancer’s incidence.Nowadays,the damage to people’s health caused by air pollution has aroused the intense concern of the Chinese society,more and more people has pay attention to the mechanism of tumorigenesis and development of lung cancer.Lung cancer is divided into squamous cell carcinoma,adenocarcinoma,gland phosphorus carcinoma,small cell carcinoma and large cell carcinoma and sarcomatoid carcinoma in the histology.In recent years,statistics show that the incidence of lung adenocarcinoma have obvious rising trend.At present,the treatment of lung cancer mainly depends on radical resection surgery,chemotherapy and radiotherapy is complementary.With the development of medical technology,molecular targeted therapy,interventional therapy and immune therapy in the treatment of lung cancer also play an increasingly important role.For lung adenocarcinoma,however,the clinical treatment effect and prognosis is worse than squamous cell carcinoma,after surgical removal of the cancer tissue,the five-year survival rate is less than 10%,distant metastasis is the leading cause of death in patients with lung adenocarcinoma.Research has shown that mitochondria may be involved in the occurrence and development of tumor.Therefore,this research adopt MTT method,immunohistochemical method,scratch test,histochemical method and Mito-tracker Green method of specific marker to investigate mitochondria’s role in the migration of lung adenocarcinoma cancer cell and A549 lung cancer cell.Methods:Both in vivo and in vitro experiment were included in this research.In vivo:Lung adenocarcinoma tissues were collected from the tumor hospital of Jilin Province,conventional paraffin embedding,sectioning(5μm)and then did immunhistochemistry of succinate dehydrogenaseIn vitro : A549 lung cancer cells were cultured by conventional method,antimycin A(mitochondrial function inhibitors),three bromine pyruvate(glycolysis functional inhibitor)and sodium pyruvate were added to the cell,the optimal drug concentrations were determined by the MTT method.Scratch test detected cell migration ability;histochemical method detect the activity of succinate dehydrogenase and lactate dehydrogenase;the mitochondrial special notation by Mito-tracker Green examined mitochondrial length changes;immunohistochemical method detected the expression of mitochondrial division protein.Results:In vivo:the result of immunohistochemical staining:the expression of succinate dehydrogenase in well-differentiated lung adenocarcinoma tissue was significantly lower than poor-differentiated lung adenocarcinoma tissue(P < 0.05);the succinate dehydrogenase expression in basal cell of well-differentiated lung adenocarcinoma was significantly lower than poor-differentiated lung adenocarcinoma basal cell(P < 0.001).In vitro:the result of MTT test showed that :the optimal concentrations of antimycin A is 150μM;the optimal concentrations of three bromine pyruvate was 200μM;the optimal concentrations of sodium pyruvate was 0.02 M.The result of scratch test showed that:there was significant difference(P<0.0001)between the cell migration distance of antimycin A group(125.6 ± 30.01μm)and control group(584.1±59.70μm);there was significant difference(P<0.001)between the cell migration distance of three bromine pyruvate group(322.4±29.16μm)and control group;there was significant difference(P<0.01)between the cell migration distance of the group of three bromine pyruvate combine with sodium pyruvate(521.1±32.92μm)and three bromine pyruvate group;there was significant difference(P<0.001)between the cell migration distance of antimycin A group and three bromine pyruvate group;there was no significant difference(P>0.05)between the cell migration distance of the group of three bromine pyruvate combine with sodium pyruvate and control group.The result of lactate dehydrogenase activity determined by histochemical test showed that there was no significant difference(P > 0.05)between the lactate dehydrogenase activity of three bromine pyruvate group(0.006± 0.003)and control group(0.008±0.004);there was no significant difference(P > 0.05)between the lactate dehydrogenase activity of the group of three bromine pyruvate combine with sodium pyruvate(0.007 ± 0.004)and three bromine pyruvate group;lactate dehydrogenase activity of antimycin A group(0.020± 0.002)was significantly higher than that of control group(P < 0.01).The result of succinate dehydrogenase activity determined by histochemical test showed that succinate dehydrogenase activity of antimycin A group(0.002 ± 0.001)was significantly higher compared with control group(0.062 ± 0.008)(P < 0.0001);there was significant difference(P < 0.0001)between the succinate dehydrogenase activity of three bromine pyruvate group(0.030 ± 0.007)and control group;there was significant difference(P < 0.0001)between the succinate dehydrogenase activity of the group of three bromine pyruvate combine with sodium pyruvate(0.057 ± 0.005)and three bromine pyruvate group;there was no significant difference(P > 0.05)between the succinate dehydrogenase activity of the group of three bromine pyruvate combine with sodium pyruvate and control group.The result of mitochondrial special notation determined by Mito-tracker Green showed that there was significant difference(P < 0.0001)between the mitochondrial length of antimycin A group(1.257±0.257μm)and control group(27.88±6.209μm);there was significant difference(P < 0.0001)between the mitochondrial length of three bromine pyruvate group(6.569 ± 2.311μm)and control group;there was no significant difference(P > 0.05)between the mitochondrial length of the group of three bromine pyruvate combine with sodium pyruvate(21.95 ± 6.661μm)and control group.The result of the expression of mitochondrial division protein determined by immunohistochemical staining showed that: there was significant difference(P < 0.0001)between the expression of mitochondrial division protein of antimycin A group(0.172 ± 0.011)and control group(0.038± 0.008);there was significant difference(P < 0.01)between the expression of mitochondrial division protein of three bromine pyruvate group(0.078 ± 0.006)and control group;there was no significant difference(P > 0.05)between the expression of mitochondrial division protein of the group of three bromine pyruvate combine with sodium pyruvate(0.047 ± 0.004)and control group.Conclution: the expression of succinate dehydrogenase was related to lung adenocarcinoma tissue differentiation degree and the lung adenocarcinoma cancer cell migration ability;mitochondrial oxidative phosphorylation and glycolysis were involved in lung adenocarcinoma cancer cell migration;glycolysis provided pyruvate for mitochondrial oxidative phosphorylation,mitochondrial oxidative phosphorylation provide enough energy for lung adenocarcinoma cancer cell migration;inhibition of mitochondrial oxidative phosphorylation enhanced mitochondrial division and reduced the lung adenocarcinoma cell migration ability. |
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