This work aimed to discover new therapeutic targets in renal clear cell carcinoma by bioinformatics and detect the effect of candidate gene TRIP13 in renal cell carcinoma(RCC)cell proliferation,migration,and invasion....This work aimed to discover new therapeutic targets in renal clear cell carcinoma by bioinformatics and detect the effect of candidate gene TRIP13 in renal cell carcinoma(RCC)cell proliferation,migration,and invasion.Differentially expressed mRNAs were screened based on The Cancer Genome Atlas(TCGA)-Kidney Renal Clear Cell Carcinoma(KIRC)databases,and functional enrichments,survival analysis,receiver operating characteristic curve(ROC),and Protein–Protein Interaction(PPI)protein interaction analysis were performed by R software to screen the candidate gene TRIP13.Then,the expression of candidate gene TRIP13 in 92 pairs of cancer and adjacent normal tissues of renal clear cell carcinoma patients were detected by qRT-PCR,western blotting,and immunochemical analysis.The TRIP13 level and clinicopathological characteristics of patients with renal clear cell carcinoma were analyzed.Using 186-O and ACHN RCC cell lines with TRIP13 overexpressing or downregulating,the effect of TRIP13 on cell viability and proliferation were detected by CCK8 and EdU staining,respectively.The migration and invasion were detected by Transwell assays.A total of 19858 differentially expressed genes,5823 differentially expressed genes,3657 up-regulated genes,and 2166 down-regulated genes were identified.TRIP13 was closed associated with cell cycle regulation,and survival and prognosis of renal clear cell carcinoma were selected as a candidate gene.The mRNA and protein levels of TRIP13 in cancer tissues were higher than that in adjacent normal tissues.TRIP13 level was significantly associated with tumor size,tumor stage,Fuhrman grade,and lymph node metastasis.TRIP13 overexpression significantly increased cell viability,proliferation,migration,and invasion,while downregulating of TRIP13 had opposite effects in both 186-O and ACHN cells.Therefore,TRIP13 promotes RCC proliferation and metastasis,which should be a novel biomarker for early diagnosis,treatment,and prognosis of RCC.展开更多
OBJECTIVE: To investigate the incidence and associated factors of multicentricity in renal cell carcinoma (RCC) in Chinese patients. METHODS: One hundred and two kidney samples from radical nephrectomy due to RCC were...OBJECTIVE: To investigate the incidence and associated factors of multicentricity in renal cell carcinoma (RCC) in Chinese patients. METHODS: One hundred and two kidney samples from radical nephrectomy due to RCC were step sectioned at 3 mm intervals and examined. All tissue abnormalities were removed, stained and examined for multicentricity. Then, on each slice of the sample, both the parenchymal margin of 15 mm beyond the pseudocapsule and tissue around the renal sinus were continuously sectioned and examined for completeness of the pseudocapsule and vascular and lymph node invasion. The relationship between muliticentricity and other pathological parameters was evaluated. RESULTS: The incidence of multicentricity was 15.7% (16/102); it was significantly lower in primary tumors 4.0 cm (4.9%, 2/41 vs 23.0%, 14/61; chi(2) = 6.055, P = 0.014). The incidence was 9.8% (8/82) in tumors without vascular invasion and 40.0% (8/20) in those with it (P = 0.003, Fisher's exact test). The incidence of multicentricity was 1.9% (1/53) in tumors with a complete pseudocapsule and 30.6% (15/49) in those without it (chi(2) = 15.885, P = 0.000). The grade, stage, subtypes and lymph node invasion of the primary tumor were not significantly associated with multicentricity. Multiple logistic regression analysis showed that pseudocapsular incompleteness and vascular invasion were two significant predictors of RCC multicentricity (P = 0.005 and 0.023). CONCLUSIONS: The incidence of multicentricity of RCC in this group of patients was in accordance with published studies. Multifocality was significantly associated with tumor size, pseudocapsule completeness and vascular invasion. NSS should be limited to tumors less than 4.0 cm when the contralateral kidney is normal and careful long-term follow-up is necessary in tumors with positive vascular invasion and incomplete pseudocapsule.展开更多
目的探讨肥大细胞对肾癌侵袭迁移能力的影响及分子机制。方法提取人肥大细胞系HMC-1的条件培养基并用其培养肾癌OSRC-2细胞,通过Transwell实验检测OSRC-2细胞体外迁移及侵袭能力的变化;采用实时定量聚合酶链反应(real time RT-PCR)及Wes...目的探讨肥大细胞对肾癌侵袭迁移能力的影响及分子机制。方法提取人肥大细胞系HMC-1的条件培养基并用其培养肾癌OSRC-2细胞,通过Transwell实验检测OSRC-2细胞体外迁移及侵袭能力的变化;采用实时定量聚合酶链反应(real time RT-PCR)及Western blot实验检测OSRC-2细胞基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)的mRNA和蛋白的表达变化;通过人TGF-β重组蛋白处理OSRC-2细胞,并通过real time RT-PCR检测MMP2 mRNA的表达变化;TGF-β信号抑制剂SB431542与HMC-1条件培养基联合处理OSRC-2细胞,通过real time RT-PCR检测OSRC-2细胞MMP2 mRNA的表达变化,通过Transwell实验检测OSRC-2细胞体外迁移及侵袭能力变化。结果HMC-1条件培养基可显著提高OSRC-2细胞的体外迁移及侵袭能力并促进MMP2表达。人TGF-β重组蛋白亦可促进OSRC-2细胞MMP2表达。加入TGF-β信号抑制剂SB431542可抑制HMC-1条件培养基引起的OSRC-2细胞MMP2表达及体外侵袭迁移。结论肥大细胞可通过分泌TGF-β上调肾癌细胞MMP2表达,进而促进侵袭转移。展开更多
目的探讨肿瘤蛋白p53基因(TP53)沉默介导PI3K/PTEN/AKT信号通路对肾透明细胞癌侵袭转移的调控机制。方法取60例肾透明细胞癌组织标本,免疫组化分析TP53蛋白在肾透明细胞癌组织中的表达;取人肾透明细胞癌细胞株RLC-310进行细胞培养;细胞...目的探讨肿瘤蛋白p53基因(TP53)沉默介导PI3K/PTEN/AKT信号通路对肾透明细胞癌侵袭转移的调控机制。方法取60例肾透明细胞癌组织标本,免疫组化分析TP53蛋白在肾透明细胞癌组织中的表达;取人肾透明细胞癌细胞株RLC-310进行细胞培养;细胞转染表达载体分为空白组(A组)、阴性对照组(B组)、sh TP53组(C组)、PTEN抑制剂bpv组(D组)、phen+sh TP53组(E组)。采用实时荧光定量PCR(qRT-PCR)和Western Blot检测各组细胞TP53、同源性磷酸酶-张力蛋白(PTEN)、磷脂酰肌醇-3-激酶(PI3K)、AKT的m RNA和蛋白表达水平;CCK-8法检测各组细胞增殖能力;划痕愈合试验检测各组细胞的迁移能力;Transwell侵袭试验检测各组细胞的侵袭能力。结果TP53蛋白在肾透明细胞癌组织中高表达(P<0.05);与A组和B组相比,C组的TP53,PI3K,AKT m RNA和蛋白表达水平显著下降,PTEN m RNA和蛋白表达水平均显著上升,且细胞增殖能力、迁移率、侵袭率均显著下降(P<0.05);D组TP53,PI3K,AKT m RNA和蛋白表达水平显著上升,PTEN m RNA和蛋白表达水平显著下降,且细胞增殖能力、迁移率、侵袭率均显著上升(P<0.05);同时,E组TP53下降(P<0.05),而其他指标与A组和B组相当(P>0.05)。结论沉默TP53基因抑制PI3K/PTEN/AKT信号通路,从而抑制肾透明细胞癌细胞浸润转移功能,并可逆转phen诱导的肾透明细胞癌细胞浸润转移。展开更多
基金supported by Grants from the Nature Science Foundation of Fujian,China(Nos.2010J01372,2015J01571).
文摘This work aimed to discover new therapeutic targets in renal clear cell carcinoma by bioinformatics and detect the effect of candidate gene TRIP13 in renal cell carcinoma(RCC)cell proliferation,migration,and invasion.Differentially expressed mRNAs were screened based on The Cancer Genome Atlas(TCGA)-Kidney Renal Clear Cell Carcinoma(KIRC)databases,and functional enrichments,survival analysis,receiver operating characteristic curve(ROC),and Protein–Protein Interaction(PPI)protein interaction analysis were performed by R software to screen the candidate gene TRIP13.Then,the expression of candidate gene TRIP13 in 92 pairs of cancer and adjacent normal tissues of renal clear cell carcinoma patients were detected by qRT-PCR,western blotting,and immunochemical analysis.The TRIP13 level and clinicopathological characteristics of patients with renal clear cell carcinoma were analyzed.Using 186-O and ACHN RCC cell lines with TRIP13 overexpressing or downregulating,the effect of TRIP13 on cell viability and proliferation were detected by CCK8 and EdU staining,respectively.The migration and invasion were detected by Transwell assays.A total of 19858 differentially expressed genes,5823 differentially expressed genes,3657 up-regulated genes,and 2166 down-regulated genes were identified.TRIP13 was closed associated with cell cycle regulation,and survival and prognosis of renal clear cell carcinoma were selected as a candidate gene.The mRNA and protein levels of TRIP13 in cancer tissues were higher than that in adjacent normal tissues.TRIP13 level was significantly associated with tumor size,tumor stage,Fuhrman grade,and lymph node metastasis.TRIP13 overexpression significantly increased cell viability,proliferation,migration,and invasion,while downregulating of TRIP13 had opposite effects in both 186-O and ACHN cells.Therefore,TRIP13 promotes RCC proliferation and metastasis,which should be a novel biomarker for early diagnosis,treatment,and prognosis of RCC.
基金ThisresearchwassupportedbyagrantfromtheLiaoningProvincialEducationalCommissionofChina (No 980 42 110 3 6)
文摘OBJECTIVE: To investigate the incidence and associated factors of multicentricity in renal cell carcinoma (RCC) in Chinese patients. METHODS: One hundred and two kidney samples from radical nephrectomy due to RCC were step sectioned at 3 mm intervals and examined. All tissue abnormalities were removed, stained and examined for multicentricity. Then, on each slice of the sample, both the parenchymal margin of 15 mm beyond the pseudocapsule and tissue around the renal sinus were continuously sectioned and examined for completeness of the pseudocapsule and vascular and lymph node invasion. The relationship between muliticentricity and other pathological parameters was evaluated. RESULTS: The incidence of multicentricity was 15.7% (16/102); it was significantly lower in primary tumors 4.0 cm (4.9%, 2/41 vs 23.0%, 14/61; chi(2) = 6.055, P = 0.014). The incidence was 9.8% (8/82) in tumors without vascular invasion and 40.0% (8/20) in those with it (P = 0.003, Fisher's exact test). The incidence of multicentricity was 1.9% (1/53) in tumors with a complete pseudocapsule and 30.6% (15/49) in those without it (chi(2) = 15.885, P = 0.000). The grade, stage, subtypes and lymph node invasion of the primary tumor were not significantly associated with multicentricity. Multiple logistic regression analysis showed that pseudocapsular incompleteness and vascular invasion were two significant predictors of RCC multicentricity (P = 0.005 and 0.023). CONCLUSIONS: The incidence of multicentricity of RCC in this group of patients was in accordance with published studies. Multifocality was significantly associated with tumor size, pseudocapsule completeness and vascular invasion. NSS should be limited to tumors less than 4.0 cm when the contralateral kidney is normal and careful long-term follow-up is necessary in tumors with positive vascular invasion and incomplete pseudocapsule.
文摘目的探讨肥大细胞对肾癌侵袭迁移能力的影响及分子机制。方法提取人肥大细胞系HMC-1的条件培养基并用其培养肾癌OSRC-2细胞,通过Transwell实验检测OSRC-2细胞体外迁移及侵袭能力的变化;采用实时定量聚合酶链反应(real time RT-PCR)及Western blot实验检测OSRC-2细胞基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)的mRNA和蛋白的表达变化;通过人TGF-β重组蛋白处理OSRC-2细胞,并通过real time RT-PCR检测MMP2 mRNA的表达变化;TGF-β信号抑制剂SB431542与HMC-1条件培养基联合处理OSRC-2细胞,通过real time RT-PCR检测OSRC-2细胞MMP2 mRNA的表达变化,通过Transwell实验检测OSRC-2细胞体外迁移及侵袭能力变化。结果HMC-1条件培养基可显著提高OSRC-2细胞的体外迁移及侵袭能力并促进MMP2表达。人TGF-β重组蛋白亦可促进OSRC-2细胞MMP2表达。加入TGF-β信号抑制剂SB431542可抑制HMC-1条件培养基引起的OSRC-2细胞MMP2表达及体外侵袭迁移。结论肥大细胞可通过分泌TGF-β上调肾癌细胞MMP2表达,进而促进侵袭转移。
文摘目的探讨肿瘤蛋白p53基因(TP53)沉默介导PI3K/PTEN/AKT信号通路对肾透明细胞癌侵袭转移的调控机制。方法取60例肾透明细胞癌组织标本,免疫组化分析TP53蛋白在肾透明细胞癌组织中的表达;取人肾透明细胞癌细胞株RLC-310进行细胞培养;细胞转染表达载体分为空白组(A组)、阴性对照组(B组)、sh TP53组(C组)、PTEN抑制剂bpv组(D组)、phen+sh TP53组(E组)。采用实时荧光定量PCR(qRT-PCR)和Western Blot检测各组细胞TP53、同源性磷酸酶-张力蛋白(PTEN)、磷脂酰肌醇-3-激酶(PI3K)、AKT的m RNA和蛋白表达水平;CCK-8法检测各组细胞增殖能力;划痕愈合试验检测各组细胞的迁移能力;Transwell侵袭试验检测各组细胞的侵袭能力。结果TP53蛋白在肾透明细胞癌组织中高表达(P<0.05);与A组和B组相比,C组的TP53,PI3K,AKT m RNA和蛋白表达水平显著下降,PTEN m RNA和蛋白表达水平均显著上升,且细胞增殖能力、迁移率、侵袭率均显著下降(P<0.05);D组TP53,PI3K,AKT m RNA和蛋白表达水平显著上升,PTEN m RNA和蛋白表达水平显著下降,且细胞增殖能力、迁移率、侵袭率均显著上升(P<0.05);同时,E组TP53下降(P<0.05),而其他指标与A组和B组相当(P>0.05)。结论沉默TP53基因抑制PI3K/PTEN/AKT信号通路,从而抑制肾透明细胞癌细胞浸润转移功能,并可逆转phen诱导的肾透明细胞癌细胞浸润转移。
