Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is asso...Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC 50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Furthermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.展开更多
Objective To observe the efficacy of deep learning(DL)model based on PET/CT and its combination with Cox proportional hazard model for predicting progressive disease(PD)of lung invasive adenocarcinoma within 5 years a...Objective To observe the efficacy of deep learning(DL)model based on PET/CT and its combination with Cox proportional hazard model for predicting progressive disease(PD)of lung invasive adenocarcinoma within 5 years after surgery.Methods The clinical,PET/CT and 5-year follow-up data of 250 patients with lung invasive adenocarcinoma were retrospectively analyzed.According to PD or not,the patients were divided into the PD group(n=71)and non-PD group(n=179).The basic data and PET/CT findings were compared between groups,among which the quantitative variables being significant different between groups were transformed to categorical variables using receiver operating characteristic(ROC)curve and corresponding cut-off value.Multivariant Cox proportional hazard model was used to select independent predicting factors of PD of lung invasive adenocarcinoma within 5 years after surgery.The patients were divided into training,validation and test sets at the ratio of 6∶2∶2,and PET/CT data in training set and validation set were used to train model and tuning parameters to build the PET/CT DL model,and the combination model was built in serial connection of DL model and the predictive factors.In test set,the efficacy of each model for predicting PD of lung invasive adenocarcinoma within 5 years after surgery was assessed and compared using the area under the curve(AUC).Results Patients'gender and smoking status,as well as the long diameter,SUV max and SUV mean of lesions measured on PET images,the long diameter,short diameter and type of lesions showed on CT were statistically different between groups(all P<0.05).Smoking(HR=1.787[1.053,3.031],P=0.031)and lesion SUV max>4.15(HR=5.249[1.062,25.945],P=0.042)were both predictors of PD of lung invasive adenocarcinoma within 5 years after surgery.In test set,the AUC of PET/CT DL model for predicting PD was 0.847,of the combination model was 0.890,of the latter was higher than of the former(P=0.036).Conclusion DL model based on PET/CT had high efficacy for predicting PD of lung invasive adenocarcinoma within 5 years after surgery.Combining with Cox proportional hazard model could further improve its predicting efficacy.展开更多
OBJECTIVE To evaluate the anticancer activity of andrographolide(AGP)and its semisynthetic analogues(SRJ09and SRJ23)in pancreatic adenocarcinoma(PDAC)cell lines harbouring therapeutically highly relevant oncogenic K-r...OBJECTIVE To evaluate the anticancer activity of andrographolide(AGP)and its semisynthetic analogues(SRJ09and SRJ23)in pancreatic adenocarcinoma(PDAC)cell lines harbouring therapeutically highly relevant oncogenic K-ras glycine-12(KRAS-G12)mutant proteins.In a landmark publication,we revealed that AGP and its derivatives bind KRAS protein to inhibit RAS signaling PNAS,110:10201-06).This discovery prompted the initiation of this investigation.METHODS The cell growth inhibitory effect of the compounds on PDAC cell lines〔PANC-1(KRAS-G12D),Capan-2(KRAS-G12V),and MIA PaCa-2(KRASG12C)〕,was assessed by MTT assay.RESULTS In comparison with AGP and SRJ09,SRJ23 showed the greatest growth inhibition in all PDAC cell lines with mutant KRAS proteins.The inhibitory effect of SRJ23 on the cell growth was similar for all PDAC cell lines.AGP exerted selective growth inhibition against PANC-1(KRAS-G12D)cells,while the growth inhibition of SRJ09 was selective towards Capan-2(KRAS-G12V)cells.CONCLUSION AGP and SRJ09 showed selectivity for PDAC cell lines with specific KRAS mutations.This suggests the mutational status of KRAS protein and the structural features of these two compounds orchestrally determined the magnitude of cell growth inhibition in PDAC cell lines.The higher potency of SRJ23 implies it could be developed into an anticancer agent for the treatment of mutant KRAS-driven malignancies.To this end,efforts are in progress to derive new molecules from this compound for further improvement of potency.展开更多
OBJECTIVE To explore the role of resistin in lung adenocarcinoma progression and its mechanism.METHODS The effect of resistin on A549 cells proliferation was detected by MTS assay.Wound-healing and transwell assays we...OBJECTIVE To explore the role of resistin in lung adenocarcinoma progression and its mechanism.METHODS The effect of resistin on A549 cells proliferation was detected by MTS assay.Wound-healing and transwell assays were used to evaluate the influence of resistin on A549 migration and invasion.Protein expression was detected by western blot.NF-k B translocation was evaluated by immunofluorescence.The expression of resistin in tumor tissue was assayed by immunohisto-chemical staining.RESULTS Compared with para-carcinoma tissues,resistin was overexpressed in tumor tissues.Resistin didn′t significantly affect A549 proliferation,but induced migration and invasion of A549.TLR4was the functional receptor of resistin in A549 cells,and resistin can bind to the second domain of TLR4.Resistin could increase p-EGFR by TLR4,induce PI3K/Akt phosphorylation and NF-k B translocation to nuclear.High resistin expression in lung adenocarcinoma tissues was correlated significantly with metastasis.Resistin was an independent predictor of overall survival.CONCLUSION Resistin promoted A549 migration and invasion by TLR4/EGFR/NF-k B pathway.Resistin was an independent prognosis predictor of lung adenocarcinoma.展开更多
Objectives:To isolate tumor metastaticsuppressing genes or relating human DNA sequencesand to study the molecular biological regulatingmechanism of tumor metastasis. Methods: HumanDNA fragments were amplified by Inter...Objectives:To isolate tumor metastaticsuppressing genes or relating human DNA sequencesand to study the molecular biological regulatingmechanism of tumor metastasis. Methods: HumanDNA fragments were amplified by Inter Alu PCRtechnique from normal human genomic DNAtransfected mouse tumor cell clones of which themetastatic phenotype had been suppressed. Thehuman origin of the amplified DNA was furtherconfirmed by PCR in situ hybridization. One展开更多
Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinom...Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.展开更多
Background and objective:Advances in high-resolution computed tomography(CT)scanning have increased the detection of small ground-glass opacity(GGO)nodules and also allowed such images to be investigated in detail.How...Background and objective:Advances in high-resolution computed tomography(CT)scanning have increased the detection of small ground-glass opacity(GGO)nodules and also allowed such images to be investigated in detail.However,it is difficult to differentiate atypical adenomatous hyperplasia(AAH)from adenocarcinoma in situ(AIS)with CT,even at follow-up,because they share many similar CT manifestations.While AAH is thought to be a precursor or even an early-stage lesion of lung adenocarcinoma,and the stepwise progression from AAH to AIS is thought to be reasonable.Therefore,the hypothesis that the attenuation of GGO is increased gradually from AAH to AIS is proposed.The aim of this study was to distinguish AAH from AIS with CT attenuation in patients with pure GGO nodules.Methods:Between January 2010 and December 2012,the CT findings in terms of the greatest diameter and mean CT attenuation(HU)were reviewed and correlated with pathology in 56 patients with AAH(n=21) and non-mucinous AIS(n=38) by two independent observers.All the 59 lesions were pure GGO nodules with size of 2 cm or smaller.To determine variability of measuring CT attenuation,we calculated the 95% confidence interval(CI)for the limits of agreement by using Bland-Altman analysis.Student t test was used to compare AAH and AIS in terms of diameter and CT attenuation.And receiver operating characteristic(ROC)curve was used to determine the optimal cut-off value of mean CT attenuation for differentiating AAH from AIS and obtain the diagnostic value.Two-tailed P value of less than 0.05 was considered to be significant.Results:For the manually measured CT attenuation,the 95% CI for the limits of agreement was-40 HU,50 HU for inter-observer variability.Although there was significant difference in nodule diameter between AAH and AIS(P=0.046),the overlap was considerable.The mean CT attenuation was (-718±53) HU(95%CI:-822,-604) for AAH,which was significantly smaller than(-600±35) HU(95%CI:-669,-531) for AIS(P=0.013).The area under curve(AUC)from ROC was 0.903 for differentiating AAH from AIS,and the cut-off value of-632 HU was optimal for differentiation between AAH and AIS,with sensitivity of 0.79,specificity of 0.95,and accuracy of 0.85.Conclusion:The mean CT attenuation can help the radiological differentiation between AAH and AIS.展开更多
文摘Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC 50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Furthermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.
文摘Objective To observe the efficacy of deep learning(DL)model based on PET/CT and its combination with Cox proportional hazard model for predicting progressive disease(PD)of lung invasive adenocarcinoma within 5 years after surgery.Methods The clinical,PET/CT and 5-year follow-up data of 250 patients with lung invasive adenocarcinoma were retrospectively analyzed.According to PD or not,the patients were divided into the PD group(n=71)and non-PD group(n=179).The basic data and PET/CT findings were compared between groups,among which the quantitative variables being significant different between groups were transformed to categorical variables using receiver operating characteristic(ROC)curve and corresponding cut-off value.Multivariant Cox proportional hazard model was used to select independent predicting factors of PD of lung invasive adenocarcinoma within 5 years after surgery.The patients were divided into training,validation and test sets at the ratio of 6∶2∶2,and PET/CT data in training set and validation set were used to train model and tuning parameters to build the PET/CT DL model,and the combination model was built in serial connection of DL model and the predictive factors.In test set,the efficacy of each model for predicting PD of lung invasive adenocarcinoma within 5 years after surgery was assessed and compared using the area under the curve(AUC).Results Patients'gender and smoking status,as well as the long diameter,SUV max and SUV mean of lesions measured on PET images,the long diameter,short diameter and type of lesions showed on CT were statistically different between groups(all P<0.05).Smoking(HR=1.787[1.053,3.031],P=0.031)and lesion SUV max>4.15(HR=5.249[1.062,25.945],P=0.042)were both predictors of PD of lung invasive adenocarcinoma within 5 years after surgery.In test set,the AUC of PET/CT DL model for predicting PD was 0.847,of the combination model was 0.890,of the latter was higher than of the former(P=0.036).Conclusion DL model based on PET/CT had high efficacy for predicting PD of lung invasive adenocarcinoma within 5 years after surgery.Combining with Cox proportional hazard model could further improve its predicting efficacy.
基金The project supported by Ministry of Education,Malaysia(Research University Grant Scheme Grant 04-02-12-2017RUFundamental Research Grant Scheme Grant 04-02-13-1324FR)
文摘OBJECTIVE To evaluate the anticancer activity of andrographolide(AGP)and its semisynthetic analogues(SRJ09and SRJ23)in pancreatic adenocarcinoma(PDAC)cell lines harbouring therapeutically highly relevant oncogenic K-ras glycine-12(KRAS-G12)mutant proteins.In a landmark publication,we revealed that AGP and its derivatives bind KRAS protein to inhibit RAS signaling PNAS,110:10201-06).This discovery prompted the initiation of this investigation.METHODS The cell growth inhibitory effect of the compounds on PDAC cell lines〔PANC-1(KRAS-G12D),Capan-2(KRAS-G12V),and MIA PaCa-2(KRASG12C)〕,was assessed by MTT assay.RESULTS In comparison with AGP and SRJ09,SRJ23 showed the greatest growth inhibition in all PDAC cell lines with mutant KRAS proteins.The inhibitory effect of SRJ23 on the cell growth was similar for all PDAC cell lines.AGP exerted selective growth inhibition against PANC-1(KRAS-G12D)cells,while the growth inhibition of SRJ09 was selective towards Capan-2(KRAS-G12V)cells.CONCLUSION AGP and SRJ09 showed selectivity for PDAC cell lines with specific KRAS mutations.This suggests the mutational status of KRAS protein and the structural features of these two compounds orchestrally determined the magnitude of cell growth inhibition in PDAC cell lines.The higher potency of SRJ23 implies it could be developed into an anticancer agent for the treatment of mutant KRAS-driven malignancies.To this end,efforts are in progress to derive new molecules from this compound for further improvement of potency.
基金The project supported by National HighTech R&D Program of China(863 Program)(2012AA02A517)National Natural Science Foundation of China(81373490,81573508,81573463)Hunan Provincial Science and Technology Plan of China(2015TP1043)
文摘OBJECTIVE To explore the role of resistin in lung adenocarcinoma progression and its mechanism.METHODS The effect of resistin on A549 cells proliferation was detected by MTS assay.Wound-healing and transwell assays were used to evaluate the influence of resistin on A549 migration and invasion.Protein expression was detected by western blot.NF-k B translocation was evaluated by immunofluorescence.The expression of resistin in tumor tissue was assayed by immunohisto-chemical staining.RESULTS Compared with para-carcinoma tissues,resistin was overexpressed in tumor tissues.Resistin didn′t significantly affect A549 proliferation,but induced migration and invasion of A549.TLR4was the functional receptor of resistin in A549 cells,and resistin can bind to the second domain of TLR4.Resistin could increase p-EGFR by TLR4,induce PI3K/Akt phosphorylation and NF-k B translocation to nuclear.High resistin expression in lung adenocarcinoma tissues was correlated significantly with metastasis.Resistin was an independent predictor of overall survival.CONCLUSION Resistin promoted A549 migration and invasion by TLR4/EGFR/NF-k B pathway.Resistin was an independent prognosis predictor of lung adenocarcinoma.
文摘Objectives:To isolate tumor metastaticsuppressing genes or relating human DNA sequencesand to study the molecular biological regulatingmechanism of tumor metastasis. Methods: HumanDNA fragments were amplified by Inter Alu PCRtechnique from normal human genomic DNAtransfected mouse tumor cell clones of which themetastatic phenotype had been suppressed. Thehuman origin of the amplified DNA was furtherconfirmed by PCR in situ hybridization. One
文摘Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.
文摘Background and objective:Advances in high-resolution computed tomography(CT)scanning have increased the detection of small ground-glass opacity(GGO)nodules and also allowed such images to be investigated in detail.However,it is difficult to differentiate atypical adenomatous hyperplasia(AAH)from adenocarcinoma in situ(AIS)with CT,even at follow-up,because they share many similar CT manifestations.While AAH is thought to be a precursor or even an early-stage lesion of lung adenocarcinoma,and the stepwise progression from AAH to AIS is thought to be reasonable.Therefore,the hypothesis that the attenuation of GGO is increased gradually from AAH to AIS is proposed.The aim of this study was to distinguish AAH from AIS with CT attenuation in patients with pure GGO nodules.Methods:Between January 2010 and December 2012,the CT findings in terms of the greatest diameter and mean CT attenuation(HU)were reviewed and correlated with pathology in 56 patients with AAH(n=21) and non-mucinous AIS(n=38) by two independent observers.All the 59 lesions were pure GGO nodules with size of 2 cm or smaller.To determine variability of measuring CT attenuation,we calculated the 95% confidence interval(CI)for the limits of agreement by using Bland-Altman analysis.Student t test was used to compare AAH and AIS in terms of diameter and CT attenuation.And receiver operating characteristic(ROC)curve was used to determine the optimal cut-off value of mean CT attenuation for differentiating AAH from AIS and obtain the diagnostic value.Two-tailed P value of less than 0.05 was considered to be significant.Results:For the manually measured CT attenuation,the 95% CI for the limits of agreement was-40 HU,50 HU for inter-observer variability.Although there was significant difference in nodule diameter between AAH and AIS(P=0.046),the overlap was considerable.The mean CT attenuation was (-718±53) HU(95%CI:-822,-604) for AAH,which was significantly smaller than(-600±35) HU(95%CI:-669,-531) for AIS(P=0.013).The area under curve(AUC)from ROC was 0.903 for differentiating AAH from AIS,and the cut-off value of-632 HU was optimal for differentiation between AAH and AIS,with sensitivity of 0.79,specificity of 0.95,and accuracy of 0.85.Conclusion:The mean CT attenuation can help the radiological differentiation between AAH and AIS.
