| Objective To investigate the expression differences of GLUT1,HK2and LDHA m RNA in lung adenocarcinoma tissues and the corresponding paracancerous tissues.To study its correlation with clinicopathological features.On this basis,explore the anti-cancer effect of Shashenmaidong Decoction on lung adenocarcinoma A549 and SPCA1 cells.Explore the effect and the mechanism of Shashenmaidong Decoction on glucose metabolism of lung adenocarcinoma.Provide scientific basis for the clinical application of Shashenmaidong Decoction in the treatment of lung adenocarcinoma.Methods 1.Specimens of lung adenocarcinoma that had recently undergone surgery were selected.Tumor and paracancerous tissue were collected from each patient.The tissues were divided into tumor group and paracancerous group.RT-q PCR was used to detect the expression of GLUT1,HK2 and LDHA m RNA in cancer tissue and corresponding paracancerous tissue.Analyze the correlation between GLUT1,HK2,LDHA and clinicopathological characteristics of patients with lung adenocarcinoma.2.The survival analysis of GLUT1,HK2 and LDHA gene expression in patients with lung adenocarcinoma was conducted by using GEPIA database.Evaluate the effect of GLUT1,HK2 and LDHA on the prognosis of patients with lung adenocarcinoma.3.Prepare medicine serum of Shashenmaidong Decoction.CCK-8method was used to determine the effect of serum containing Shashenmaidong Decoction on the proliferation of human lung adenocarcinoma A549 and SPCA1 cells.Apoptosis of human lung adenocarcinoma A549 and SPCA1 cells was detected by Annexin V-FITC flow cytometry double staining.4.Human lung adenocarcinoma A549 and SPCA1 cells were cultured in vitro.After 48h intervention with drug-containing serum of Shashenmaidong Decoction,RT-q PCR and Western blot were used to detect the effects of drug-containing serum of Shashenmaidong Decoction on the expression levels of GLUT1,HC2,LDHA and HIF-1?genes and protein levels.5.Cell scratch test and Transwell invasion test were used to evaluate the effects of drug-containing serum of Shashen maidong Decoction on the migration and invasion of human lung adenocarcinoma A549 and SPCA1 cells.6.The effects of drug-containing serum of Shashenmaidong Decoction on lactic acid production and glucose consumption of human lung adenocarcinoma cells were detected by glucose detection kit and lactic acid detection kit.Results 1.The positive expression rate of GLUT1 in 36 lung adenocarcinoma tissues was 86.11%(31/36),and in the corresponding paracancerous tissues,the positive expression rate of GLUT1 was27.78%(10/36),thedifferencewasstatistically significant(c~2=24.982,P=0.000),and the expression level of GLUT1m RNA in 36 lung adenocarcinoma tissues was significantly higher than that in paracancerous normal tissues,the difference was statistically significant(P<0.01).2.The positive expression rate of HK2 in the 36 lung adenocarcinoma tissues was 75.00%(27/36),and in the corresponding paracancerous tissues,the positive expression rate of HK2 was44.44%(16/36),and the difference between the two was statistically significant(c~2=6.986,P=0.005).The expression of HK2 m RNA in 36lung adenocarcinoma tissues was significantly higher than that in adjacent normal tissues,and the difference was statistically significant(P<0.01).3.The positive expression rate of LDHA in 36 lung adenocarcinoma tissues was 80.56%(29/36),and in the corresponding paracancerous tissues,the positive expression rate of LDHA was33.33%(12/36),the difference between the two was statistically significant(c~2=14.762,P=0.000).The expression of LDHA m RNA in36 lung adenocarcinoma tissues was significantly higher than that in adjacent tissues,the difference was statistically significant(P<0.01).4.There were no significant differences in the expression of GLUT1,HK2 and LDHA in lung adenocarcinoma tissues with different gender,age,lymph node metastasis or stage(P>0.05).5.The GEPIA database was used to analyze 478 patients with lung adenocarcinoma.The results showed that the patients with high expression of GLUT1,HK2 and LDHA had poor prognosis,and the difference was statistically significant(P<0.05).6.CCK-8 method and Annexin V-fitc flow double staining method were used to detect the effects of drug-containing serum of Shachenmaidong Decoction on the proliferation and apoptosis of human lung adenocarcinoma cells.The results showed that the drug-containing serum of Shachenmaidong Decoction could inhibit the proliferation of human lung adenocarcinoma A549 and SPCA1 cells,and promote their apoptosis.7.According to the results of RT-q PCR and Western blot,drug-containing serum of Shashenmaidong Decoction can cut GLUT1,HK2,LDHA m RNA level and protein expression,further studies have found that Shashenmaidong Decoction can also lower the expression of HIF–1?,show that Shashenmaidong Decoction may influence sugar metabolism process through the HIF-1?signaling pathway and achieve the purpose of anti-tumor.8.The results of cell scratch test and Transwell invasion test showed that the drug-containing serum of Shashen Maidong Decoction could significantly inhibit the migration and invasion of human lung adenocarcinoma A549 and SPCA1 cells.9.Lactic acid and glucose test results showed that compared with the control group,Shashenmaidong decoction group could inhibit the glucose consumption and lactic acid production of human lung adenocarcinoma cells,with statistical significance(P<0.05).Conclusion 1.The expressions of GLUT1,HK2 and LDHA in lung adenocarcinoma tissues were higher than those in adjacent tissues.However,gender,age,lymph node metastasis and tumor stage of lung adenocarcinoma patients were not correlated.2.GLUT1,HK2,and LDHA may be involved in the occurrence and progression of lung adenocarcinoma,and the high expression of GLUT1,HK2,and LDHA is associated with poorer prognosis of patients.3.Shashenmaidong Decoction has a good anti-lung adenocarcinoma effect,and its anti-lung adenocarcinoma mechanism may be related to the regulation of glucose metabolism level of tumor cells. |
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