p53免疫组化与TP53突变状态之间的关系一直存在争议。本组研究重新评估p53免疫组化提示TP53突变状态的价值。通过外显子测序,共检测了157例弥漫型胶质瘤（ WHO Ⅱ～Ⅳ级）冷冻组织的TP53基因第4～10外显子。对应所有病例的福尔马林固定...p53免疫组化与TP53突变状态之间的关系一直存在争议。本组研究重新评估p53免疫组化提示TP53突变状态的价值。通过外显子测序,共检测了157例弥漫型胶质瘤（ WHO Ⅱ～Ⅳ级）冷冻组织的TP53基因第4～10外显子。对应所有病例的福尔马林固定石蜡包埋组织检测p53免疫组化（克隆号DO-7）,评估表达强度和比率。66例突变病例中发现72个突变,包括60个错义突变,5个无义突变,4个缺失和3个剪切位点变异。通过 ROC曲线分析发现大于10%的肿瘤细胞强阳性可最大程度地提示突变。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
Background/Aims: Hepatic undifferentiated (embryonal) sarcoma (HUS) is an exc eptional hepatic malignant tumor in adults. Genetic studies were never reported in adult cases. Methods: In this study concerning three cas...Background/Aims: Hepatic undifferentiated (embryonal) sarcoma (HUS) is an exc eptional hepatic malignant tumor in adults. Genetic studies were never reported in adult cases. Methods: In this study concerning three cases of HUS occurring i n adult, we studied the three classical ways of carcinogenesis i.e. the TP53 (p5 3), Wnt (CTNNB1/β - catenin and AXIN1) and telomerase (hTERT) pathways. We stu died the expression of p53, β - catenin and telomerase catalytic subunit hTERT by immunohistochemistry in the three cases; we determined TP53 gene mutation in two cases and the genome- wide allelotype, AXIN1, and CTNNB1/β - catenin gen e mutation in one case. Results: Immunohistochemistry showed an overexpression o f p53 in more than 80% of tumoral cells; furthermore, mutations of TP53 were o bserved in two cases, involving the sequence- specific DNA binding domain. In c ontrast, no mutation was found in CTNNB1/β - catenin and AXIN1 genes. Tumoral cells did not show hTERT staining nor nuclear expression of β - catenin. In ad dition, allelotype analysis in one case showed loss of heterozygosity of chromos ome 7p, 11p, 17p, 22q, and allelic imbalance of 1p, 8p, 20q. Conclusions: In thi s report of HUS in three adult patients, we emphasize the role of TP53 pathway i n carcinogenesis of this rare tumor. This point could be of interest for therape utic strategies.展开更多
Background: Basal cell carcinoma (BCC) of the skin is the most common human cancer. The genetic alterations underlying BCC development are only partly understood. Objectives: To investigate further the molecular genet...Background: Basal cell carcinoma (BCC) of the skin is the most common human cancer. The genetic alterations underlying BCC development are only partly understood. Objectives: To investigate further the molecular genetics of sporadic BCCs, we performed mutation analyses of 10 skin cancer-associated genes in 42 tumours. Methods: Single-strand conformational polymorphism analysis followed by DNA sequencing was used to screen for mutations in the sonic hedgehog pathway genes PTCH, SMOH, SUFUH and GLI1, in the TP53 tumour suppressor gene, and in the proto-oncogenes NRAS, KRAS, HRAS, BRAF and CTNNB1. Microsatellite markers flanking the PTCH, SUFUH and TP53 loci at 9q22, 10q24 and 17p13, respectively, were studied for loss of heterozygosity (LOH). Results: PTCH mutations were found in 28 of 42 tumours (67% ). Microsatellite analysis revealed LOH on 9q22 in 20 of 38 tumours investigated (53% ), including 14 tumours with and six tumours without PTCH mutations. SMOH mutations were identified in four of the 42 BCCs (10% ) while two tumours demonstrated mutations in SUFUH, including one missense mutation and one silent mutation. None of the BCCs showed LOH at markers flanking the SUFUH locus. Seventeen BCCs (40% ) carried TP53 mutations, with only three tumours showing evidence of biallelic TP53 inactivation. TP53 mutations were present in BCCs with and without mutations in PTCH, SMOH or SUFUH. Interestingly, 72% of the TP53 alterations were presumably ultraviolet (UV)- induced transition mutations. In contrast, only 40% of the PTCH and SMOH alterations corresponded to UV signature mutations. No mutations were identified in GLI1, NRAS, KRAS, HRAS, BRAF or CTNNB1. Conclusions: Our data confirm the importance of PTCH, SMOH and TP53 mutations in the pathogenesis of sporadic BCCs. SUFUH alterations are restricted to individual cases while the other investigated genes do not appear to be important targets for mutations in BCCs.展开更多
文摘p53免疫组化与TP53突变状态之间的关系一直存在争议。本组研究重新评估p53免疫组化提示TP53突变状态的价值。通过外显子测序,共检测了157例弥漫型胶质瘤（ WHO Ⅱ～Ⅳ级）冷冻组织的TP53基因第4～10外显子。对应所有病例的福尔马林固定石蜡包埋组织检测p53免疫组化（克隆号DO-7）,评估表达强度和比率。66例突变病例中发现72个突变,包括60个错义突变,5个无义突变,4个缺失和3个剪切位点变异。通过 ROC曲线分析发现大于10%的肿瘤细胞强阳性可最大程度地提示突变。
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。
文摘Background/Aims: Hepatic undifferentiated (embryonal) sarcoma (HUS) is an exc eptional hepatic malignant tumor in adults. Genetic studies were never reported in adult cases. Methods: In this study concerning three cases of HUS occurring i n adult, we studied the three classical ways of carcinogenesis i.e. the TP53 (p5 3), Wnt (CTNNB1/β - catenin and AXIN1) and telomerase (hTERT) pathways. We stu died the expression of p53, β - catenin and telomerase catalytic subunit hTERT by immunohistochemistry in the three cases; we determined TP53 gene mutation in two cases and the genome- wide allelotype, AXIN1, and CTNNB1/β - catenin gen e mutation in one case. Results: Immunohistochemistry showed an overexpression o f p53 in more than 80% of tumoral cells; furthermore, mutations of TP53 were o bserved in two cases, involving the sequence- specific DNA binding domain. In c ontrast, no mutation was found in CTNNB1/β - catenin and AXIN1 genes. Tumoral cells did not show hTERT staining nor nuclear expression of β - catenin. In ad dition, allelotype analysis in one case showed loss of heterozygosity of chromos ome 7p, 11p, 17p, 22q, and allelic imbalance of 1p, 8p, 20q. Conclusions: In thi s report of HUS in three adult patients, we emphasize the role of TP53 pathway i n carcinogenesis of this rare tumor. This point could be of interest for therape utic strategies.
文摘Background: Basal cell carcinoma (BCC) of the skin is the most common human cancer. The genetic alterations underlying BCC development are only partly understood. Objectives: To investigate further the molecular genetics of sporadic BCCs, we performed mutation analyses of 10 skin cancer-associated genes in 42 tumours. Methods: Single-strand conformational polymorphism analysis followed by DNA sequencing was used to screen for mutations in the sonic hedgehog pathway genes PTCH, SMOH, SUFUH and GLI1, in the TP53 tumour suppressor gene, and in the proto-oncogenes NRAS, KRAS, HRAS, BRAF and CTNNB1. Microsatellite markers flanking the PTCH, SUFUH and TP53 loci at 9q22, 10q24 and 17p13, respectively, were studied for loss of heterozygosity (LOH). Results: PTCH mutations were found in 28 of 42 tumours (67% ). Microsatellite analysis revealed LOH on 9q22 in 20 of 38 tumours investigated (53% ), including 14 tumours with and six tumours without PTCH mutations. SMOH mutations were identified in four of the 42 BCCs (10% ) while two tumours demonstrated mutations in SUFUH, including one missense mutation and one silent mutation. None of the BCCs showed LOH at markers flanking the SUFUH locus. Seventeen BCCs (40% ) carried TP53 mutations, with only three tumours showing evidence of biallelic TP53 inactivation. TP53 mutations were present in BCCs with and without mutations in PTCH, SMOH or SUFUH. Interestingly, 72% of the TP53 alterations were presumably ultraviolet (UV)- induced transition mutations. In contrast, only 40% of the PTCH and SMOH alterations corresponded to UV signature mutations. No mutations were identified in GLI1, NRAS, KRAS, HRAS, BRAF or CTNNB1. Conclusions: Our data confirm the importance of PTCH, SMOH and TP53 mutations in the pathogenesis of sporadic BCCs. SUFUH alterations are restricted to individual cases while the other investigated genes do not appear to be important targets for mutations in BCCs.
