Objective To study the relationship between the expression of long non-coding RNA lung adenocarcinoma metastasis-associated transcript 1(lncRNA MALAT1)and microRNA(miR)-181a-5p in lung adenocarcinoma tissues and the signal pathway of Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3),clinicopathological features and prognosis.Methods 218 patients with lung adenocarcinoma who had surgical resection at our institution between January 2018 and May 2021 had their cancer tissues and nearby normal lung tissues collected,the levels of lncRNA MALAT1,miR-181a-5p and key factors of JAK2/STAT3 signaling pathway(JAK2 mRNA,STAT3 mRNA)in lung adenocarcinoma tissues and adjacent tissues were detected by reverse transcription polymerase chain reaction(RT-PCR).The correlation between the expression levels of lncRNA MALAT1 and miR-181a-5p in cancer tissues of lung adenocarcinoma patients and the levels of key factors in JAK2/STAT3 signaling pathway were analyzed by Pearson test.The relationship between the expression levels of lncRNA MALAT1 and miR-181a-5p and the clinicopathological features of lung adenocarcinoma patients were analyzed.Patients with lung adenocarcinoma were followed up for 3 years,and their prognosis was counted,the 3-year overall survival rate of lncRNA MALAT1 and miR-181a-5p low/high expression groups were analyzed by Kaplan-Meier method.The prognostic factors were analyzed by univariate and multivariate COX risk proportional regression models.Results In lung adenocarcinoma tissues,the expression levels of lncRNA MALAT1,JAK2,and STAT3 mRNA were substantially greater(P＜0.05)than in neighboring normal lung tissues,whereas the expression level of miR-181a-5p was significantly lower(P＜0.05)in compared to nearby normal lung tissues.Pearson test results showed that,lncRNA MALAT1 was positively correlated with JAK2 and STAT3 mRNA expression levels in cancer tissues of patients with lung adenocarcinoma(P＜0.05,r=0.526、0.483),and miR-181a-5p was negatively correlated with JAK2 and STAT3 mRNA expression levels in cancer tissues of patients with lung adenocarcinoma(P＜0.05,r=-0.430、-0.493).lncRNA MALAT1 had a considerably greater expression rate and miR-181a-5p had a significantly lower expression rate in patients with TNM stage Ⅲa,lymph node metastasis and poorly differentiated lung adenocarcinoma than in patients with TNM stage Ⅰ-Ⅱ,without lymph node metastasis and moderately well differentiated lung adenocarcinoma(P＜0.05).Three patients were lost during the 3-year follow-up of 218 patients with lung adenocarcinoma,and the 3-year overall survival rate was 58.14％(125/215).The 3-year overall survival rate of the lncRNA MALAT1 high expression group was considerably lower than that of the lncRNA MALAT1 low expression group.The miR-181a-5p high expression group had a substantially greater(P＜0.05).Lymph node metastasis,TNM stage Ⅲ a,decreased expression level of miR-181a-5p,and increased expression level of lncRNA MALAT1 are risk factors for the prognosis of patients with lung adenocarcinoma(P＜0.05).Conclusion The low expression of miR-181a-5p and the high expression of lncRNA MALAT1 in lung adenocarcinoma tissues are related to TNM stage Ⅲa,lymph node metastasis and poor prognosis,which may promote the progression of lung adenocarcinoma and cause poor prognosis by activating JAK2/STAT3 signaling pathway.

Objective To investigate the effect of forsythigenin on the malignant progression of lung cancer cells by regulating the cyclic adenosine monophosphate/exchange protein directly activated by cAMP1/Ras-associated protein 1(cAMP/EPAC1/RAP1)signaling pathway.Methods Lung cancer cell line A549 was cultured in vitro and grouped into the control group,the low dose forsythigenin group(25 mg/L),the medium dose forsythigenin group(50 mg/L),the high dose forsythigenin group(100 mg/L),the high dose forsythigenin+specific increase in intracellular cAMP content(pertussis toxin PTX)group(100 mg/L forsythigenin+5 μmol/L PTX)and high dose forsythigenin+EPAC1 antagonist(ESI-09)group(100 mg/L forsythigenin+1.5 μmol/L ESI-09).CCK-8 experiment was applied to detect cell proliferation.Scratch test was applied to detect cell migration.Flow cytometry was applied to detect cell apoptosis.Transwell was applied to detect cell invasion.ELISA method was applied to detect cAMP level in cell supernatant.Western blot assay was applied to detect expression levels of cAMP/EPAC1/RAP1 signaling pathway proteins and apoptotic proteins[B lymphoblastoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)].Results Compared with the control group,the OD450 value of A549 cells,number of cell invasions,scratch healing rate,level of cAMP,expression levels of Bcl-2,EPAC1 and RAP1 proteins were significantly reduced in the low dose,medium dose and high dose forsythigenin groups,and the expression of Bax protein and the rate of cell apoptosis were significantly increased in a dose-dependent manner(P＜0.05).Compared with the high-dose forsythigenin group,the OD450 value of A549 cells,scratch healing rate,number of cell invasions,level of cAMP,expression levels of Bcl-2,EPAC1 and RAP1 proteins were obviously increased in the high-dose forsythigenin+PTX group,the expression of Bax protein and the apoptosis rate were obviously reduced(P＜0.05).Levels of all indexes in the high dose forsythigenin+ESI-09 group were opposite.Conclusion Forsythigenin inhibits proliferation,migration,and invasion of A549 cells and promotes apoptosis by down-regulating the cAMP/EPAC1/RAP1 signaling pathway.

Objective To study the relationship between the expression of long non-coding RNA lung adenocarcinoma metastasis-associated transcript 1(lncRNA MALAT1)and microRNA(miR)-181a-5p in lung adenocarcinoma tissues and the signal pathway of Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3),clinicopathological features and prognosis.Methods 218 patients with lung adenocarcinoma who had surgical resection at our institution between January 2018 and May 2021 had their cancer tissues and nearby normal lung tissues collected,the levels of lncRNA MALAT1,miR-181a-5p and key factors of JAK2/STAT3 signaling pathway(JAK2 mRNA,STAT3 mRNA)in lung adenocarcinoma tissues and adjacent tissues were detected by reverse transcription polymerase chain reaction(RT-PCR).The correlation between the expression levels of lncRNA MALAT1 and miR-181a-5p in cancer tissues of lung adenocarcinoma patients and the levels of key factors in JAK2/STAT3 signaling pathway were analyzed by Pearson test.The relationship between the expression levels of lncRNA MALAT1 and miR-181a-5p and the clinicopathological features of lung adenocarcinoma patients were analyzed.Patients with lung adenocarcinoma were followed up for 3 years,and their prognosis was counted,the 3-year overall survival rate of lncRNA MALAT1 and miR-181a-5p low/high expression groups were analyzed by Kaplan-Meier method.The prognostic factors were analyzed by univariate and multivariate COX risk proportional regression models.Results In lung adenocarcinoma tissues,the expression levels of lncRNA MALAT1,JAK2,and STAT3 mRNA were substantially greater(P＜0.05)than in neighboring normal lung tissues,whereas the expression level of miR-181a-5p was significantly lower(P＜0.05)in compared to nearby normal lung tissues.Pearson test results showed that,lncRNA MALAT1 was positively correlated with JAK2 and STAT3 mRNA expression levels in cancer tissues of patients with lung adenocarcinoma(P＜0.05,r=0.526、0.483),and miR-181a-5p was negatively correlated with JAK2 and STAT3 mRNA expression levels in cancer tissues of patients with lung adenocarcinoma(P＜0.05,r=-0.430、-0.493).lncRNA MALAT1 had a considerably greater expression rate and miR-181a-5p had a significantly lower expression rate in patients with TNM stage Ⅲa,lymph node metastasis and poorly differentiated lung adenocarcinoma than in patients with TNM stage Ⅰ-Ⅱ,without lymph node metastasis and moderately well differentiated lung adenocarcinoma(P＜0.05).Three patients were lost during the 3-year follow-up of 218 patients with lung adenocarcinoma,and the 3-year overall survival rate was 58.14％(125/215).The 3-year overall survival rate of the lncRNA MALAT1 high expression group was considerably lower than that of the lncRNA MALAT1 low expression group.The miR-181a-5p high expression group had a substantially greater(P＜0.05).Lymph node metastasis,TNM stage Ⅲ a,decreased expression level of miR-181a-5p,and increased expression level of lncRNA MALAT1 are risk factors for the prognosis of patients with lung adenocarcinoma(P＜0.05).Conclusion The low expression of miR-181a-5p and the high expression of lncRNA MALAT1 in lung adenocarcinoma tissues are related to TNM stage Ⅲa,lymph node metastasis and poor prognosis,which may promote the progression of lung adenocarcinoma and cause poor prognosis by activating JAK2/STAT3 signaling pathway.

Objective To investigate the effect of forsythigenin on the malignant progression of lung cancer cells by regulating the cyclic adenosine monophosphate/exchange protein directly activated by cAMP1/Ras-associated protein 1(cAMP/EPAC1/RAP1)signaling pathway.Methods Lung cancer cell line A549 was cultured in vitro and grouped into the control group,the low dose forsythigenin group(25 mg/L),the medium dose forsythigenin group(50 mg/L),the high dose forsythigenin group(100 mg/L),the high dose forsythigenin+specific increase in intracellular cAMP content(pertussis toxin PTX)group(100 mg/L forsythigenin+5 μmol/L PTX)and high dose forsythigenin+EPAC1 antagonist(ESI-09)group(100 mg/L forsythigenin+1.5 μmol/L ESI-09).CCK-8 experiment was applied to detect cell proliferation.Scratch test was applied to detect cell migration.Flow cytometry was applied to detect cell apoptosis.Transwell was applied to detect cell invasion.ELISA method was applied to detect cAMP level in cell supernatant.Western blot assay was applied to detect expression levels of cAMP/EPAC1/RAP1 signaling pathway proteins and apoptotic proteins[B lymphoblastoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)].Results Compared with the control group,the OD450 value of A549 cells,number of cell invasions,scratch healing rate,level of cAMP,expression levels of Bcl-2,EPAC1 and RAP1 proteins were significantly reduced in the low dose,medium dose and high dose forsythigenin groups,and the expression of Bax protein and the rate of cell apoptosis were significantly increased in a dose-dependent manner(P＜0.05).Compared with the high-dose forsythigenin group,the OD450 value of A549 cells,scratch healing rate,number of cell invasions,level of cAMP,expression levels of Bcl-2,EPAC1 and RAP1 proteins were obviously increased in the high-dose forsythigenin+PTX group,the expression of Bax protein and the apoptosis rate were obviously reduced(P＜0.05).Levels of all indexes in the high dose forsythigenin+ESI-09 group were opposite.Conclusion Forsythigenin inhibits proliferation,migration,and invasion of A549 cells and promotes apoptosis by down-regulating the cAMP/EPAC1/RAP1 signaling pathway.

AIM:To investigate the effects of grasp seed procyanidins(GSP) on homocysteine-induced proliferation and migration in vascular smooth muscle cells(VSMC) and related molecular mechanisms.METHODS: Cell count and -TdR assay were used for detecting cell proliferation and DNA synthesis,ELISA assay was used for detecting inflammatory response,DCFH-DA assay for examining the levels of reactive oxygen species(ROS),Western blotting for detecting protein expression.RESULTS: Homocysteine(0.1-1 mmol/L) increased VSMC proliferation and migration,and the levels of ROS were in a dose-dependent manner.The results of Western blotting showed that homocysteine significantly increased the expression of MCP-1,IL-6 and TNF-?.However,Compared with control group,in GSP(5-20 g/L) group,the increased VSMC proliferation,migration and the production of ROS and the expression of MCP-1,IL-6 and TNF-? mediated by homocysteine were markedly suppressed.EMSA showed that in GSP treatment group,the NF-?B activation was also almost completely inhibited.CONCLUSION: GSP inhibits homocysteine-induced VSMC proliferation,migration and inflammatory response through interfering with ROS dependent on NF-?B signal pathway.