| Objective:To analyze the relationship between lactate dehydrogenase A(LDHA)and serum LDH,SUVmax,distant metastasis and prognosis in de novo lung adenocarcinoma through clinical data and resected tumor tissues.Futher to investigate the role of LDHA in the energy metabolism and various biological behaviors in A549 cells,and explore the possible regulatory pathway.Methods:1.(1)The patients who involved according to the inclusion and exclusion criteria were collected the clinical characteristics,TNM staging,levels of serum LDH and tumor markers(TMs).Age,gender,smoking history,the status of EGFR,T stage and N stage were matched by propensity scores,then the screened patients were divided into Distant metastasis group(DG)and Control group(CG)through M stage.The difference of serum LDH and TMs between two groups were explored.(2)All involved individuals were followed up except non-neoplastic death,secondary primary tumor,incomplete folloe-up data.The final cases were divided into Normal group(NG)and Elevated group(EG)by the levels of serum LDH and TMs.Overall survival(OS)were compared between two groups.2.The lung adenocarcinomas who underwent PET/CT befor anti-cancer treatment were involved by the inclusion and exclusion criteria.Clinical data,such as gender,age,smoking status,TNM staging were recorded.6 cases who underwent surgery were selected to detect and compare the expression of LDHA in tumor and para-tumor tissues.Furthermore,the association between the expreesion of tumor tissue LDHA,serum LDH concentrtion and SUVmax value were explored.Besides,the relationship between the level of serum LDH and SUVmax value were detected in the divided groups according to median serum LDH concentration and SUVmax value in the another patients who underwent PET/CT.3.Cell experiments:(1)Transient transfected A549 cells,then screened and identified the optimal plasmid of over-expression LDHA,interfering LDHA and interfering PTEN.As a specific inhibitor of mTOR,Rapamycin were used to down-regulate mTOR and the proper concentration were determined.(2)A549 cells were divided into eight groups: A549 control group(AC),Negative control group(NC),LDHA over-expression group(LU),Silencing control group(SC),LDHA interference group(LD),Rapamycin group(RAP),PTEN interference group(PD)and PTEN interference+LDHA interference group(PD+LD).The glucose consumption,lactate production,ATP production,proliferation,migration,invasion,cell cycle and apoptosis of each group were detected.Results:1.(1)1240 patients were selected and 620 in each group after propensity score matching.Different with the general information,the levels of LDH and TMs were higher in MG than CG(all P<0.05).Logistic regression showed high LDH(OR=1.598,95%CI: 1.169~2.184,P=0.003),high CEA(OR=1.614,95%CI: 1.245~2.092,P<0.001)、high CYFRA211(R=1.640,95%CI: 1.237~2.175,P=0.001)and high CA125(OR=2.668,95%CI: 2.020~3.524,P<0.001)were associated with distant metastasis.(2)A total of 1102 patients were finally followed up.The median time of follow-up was 26.63 months with 95%CI: 24.26~29.01 months,and the median OS was 20.33 months with 95%CI: 18.85~21.81 months.The median OS in the LDH NG was higher than that in the LDH EG(24.53 months vs 13.93 months,P<0.001).2.The expression of LDHA in tumor tissue was higher than that in para-tumor tissue(P<0.05),and positive related to serum LDH concentration and SUVmax value.Among another 280 patients,the individuals with lymph node metastasis,distant metastasis and SUVmax higher than 9.0 gain evaluated srum LDH concentration(all P<0.05).In addition,cases with over 60 years of age,female,lymph node and distant metastasis acquired increased SUVmax value(all P<0.05).What’s more,serum LDH was positively correlated with SUVmax value(r=0.187,P=0.002)).3.(1)Energy metabolism: The glucose consumption,lactate and ATP production were increased in LU and PD(all P<0.01).On the contrary,the glucose consumption,lactate and ATP production were decreased in LD and RAP(all P<0.01).Addtiongally,the glucose consumption,lactate and ATP production in PD+LD were lower than PD(all P<0.05),but the same as LD(P=0.150/0.694/0.168).(2)Bilogical behavior: Cell proliferation,migration and invasion were enhanced in LU and PD(all P<0.01)and weakened in LD and RAP(all P<0.01).Cell proliferation,migration and invasion in PD+LD were abated than that in PD(all P<0.05),but intensified than that in LD(all P<0.01)except cell invasion(P=0.467).(3)Cell cycle and apoptosis: Phase S cells decreased but phase G2 cells increased in LU and PD(all P<0.05).Differently,the number of phase G1 cells raised and that of phase G2 cells descend in LD,RAP and PD+LD(all P<0.05).The percentage of apoptosis stay still in LU and PD(all P>0.05),but elevated in LD and RAP(all P<0.01).By the way,the apoptosis rate of PD+LD was between PD and LD(all P<0.01).Conclusion:1.The patients with elevated level of serum LDH gain higher risk of distant metastasis and poor prognosis compared with cases with normal level of serum LDH.2.The expression of LDHA in tumor tissue was higher than that in para-tumor tissue and associated with serum LDH concentration and SUVmax value.The level of serum LDH was positive correlated with SUVmax value.3.In cell experiment,the glucose consumption,lactate and ATP produciont were increased when LDHA and mTOR were up-regulated,cell proliferation,migration and invasion were also enhanced.On the contrary,enegy metabolism,cell proliferation,migration and invasion were all wakened,and apoptosis was enhanced when LDHA and mTOR were down-regulated.What’s more,the glucose metabolism which mTOR was up-regulated but LDHA was down-regulated decreased to that of LDHA down-regulation.However,cell profiration,migration,invasion,apoptosis and cell cycle were stay between mTOR up-regulation only and LDHA down-regulation only.It provide an idea for molecular target selection. |
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