Hh信号通路在口腔鳞状细胞癌中的激活及意义

目的： 研究口腔鳞状细胞癌中Shh、Ptch、Gli-1在基因及蛋白水平的表达,并探讨其阳性表达与口腔鳞状细胞癌生物学特性之间的关系。方法：采用免疫组织化学方法分别检测40例口腔鳞癌和30例正常口腔黏膜中Shh、Ptch、Gli-1蛋白的表达;RT-PCR法检测10例口腔鳞癌组织及5例正常口腔黏膜组织中Shh、Gli-1和Ptch mRNA的表达。结果：免疫组织化学结果显示,Shh、Ptch和Gli-1蛋白在口腔鳞癌中阳性表达率分别为62.5%、60.0％和65.0％,在正常口腔黏膜中3种蛋白均不表达。非参数统计分析示,Shh及Gli-1的阳性表达与肿瘤的大小、淋巴结转移及临床分期相关 (P<0.05),Ptch蛋白的表达与淋巴结转移呈显著正相关 (P<0.05)。RT-PCR结果显示,Shh、Ptch、Gli-1 mRNA在口腔鳞癌组织中的阳性表达分别为60%、50%、70%,明显高于正常口腔黏膜,差异有统计学意义 (P<0.05)。结论：Hh信号通路在口腔鳞状细胞癌组织中被激活并与口腔鳞癌的发生、发展、转移关系密切。     

Hedgehog信号通路相关蛋白在头颈鳞状细胞癌中的表达及临床意义

目的:探讨头颈鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)中Hedgehog信号通路成员Shh、Ptch1和Gli1蛋白表达的关系及其意义。方法:利用生物信息学方法分析Shh、Ptch1、Gli1在HNSCC和正常组织中的表达差异预测本研究可行性,采用免疫组化法检测84例头颈鳞状细胞癌及10例正常黏膜上皮组织中Shh、Ptch1和Gli1蛋白表达,分析Shh、Ptch1和Gli1蛋白表达水平与头颈鳞状细胞癌患者临床病理特征的关系。结果:头颈鳞状细胞癌组织中Shh、Ptch1和Gli1蛋白呈高表达（分别为65.5%、60.7%和63.1%）,均高于正常黏膜上皮组织（分别为20%、30%和30%）。Shh蛋白表达与头颈鳞状细胞癌患者临床分期和淋巴结转移有关(P<0.05),而与患者年龄、性别、肿瘤局部侵袭和远处转移均无关(P>0.05);Ptch1蛋白表达与头颈鳞状细胞癌患者临床病理特征均无关(P>0.05);Gli1蛋白表达仅与头颈鳞状细胞癌患者淋巴结转移有关(P<0.05)。Shh、Ptch1和Gli1蛋白的表达水平在鼻咽鳞癌和口咽鳞癌中没有差异。Shh与Ptch1、Gli1蛋白表达呈正相关。结论:Shh、Ptch1和Gli1蛋白在头颈鳞状细胞癌中高表达,与头颈鳞状细胞癌的发生、发展密切相关,是头颈鳞状细胞癌防治的一个重要靶标。     

口腔鳞状细胞癌中MMP9、CD44v6表达及其意义

[目的]研究基质金属蛋白酶-9（matrix metalloproteinase-9,MMP9）、CD44拼接变异体6（CD44 splice variant 6,CD44v6）在口腔鳞状细胞癌（OSCC）中的表达及其相互关系.[方法]应用S-P免疫组化法检测82例口腔鳞状细胞癌,40例口腔黏膜鳞状上皮非典型增生和30例正常口腔黏膜中MMP9和CD44v6的表达.[结果]（1）MMP9在OSCC组中的阳性率（84.1%）显著高于非典型增生组（17.5%）和正常口腔黏膜组（6.6%）.MMP9的阳性表达率与淋巴结转移、肿瘤分化程度相关.（2）CD44v6在OSCC组中的阳性率（78.4%）显著高于非典型增生组（30.0%）和正常口腔黏膜（13.3%）.CD44v6的高表达率与淋巴结转移相关.（3）OSCC中MMP9的表达与CD44v6表达呈正相关.[结论]MMP9、CD44v6对OSCC发生过程具有重要意义,MMP9和CD44v6的表达在OSCC淋巴结转移过程中可能呈协同作用.     

半乳糖凝集素-9和T细胞免疫球蛋白黏蛋白分子3在口腔扁平苔藓和口腔鳞状细胞癌组织中的表达及临床意义

目的探讨半乳糖凝集素-9(Galectin-9)和T细胞免疫球蛋白黏蛋白分子3(TIM3)在口腔扁平苔藓和口腔鳞状细胞癌组织中的表达及临床意义。方法收集42例口腔扁平苔藓组织、25例口腔鳞状细胞癌组织及30例正常口腔黏膜组织,采用免疫组织化学染色法检测3种组织中Galectin-9和TIM3的表达情况,分析Galectin-9和TIM3表达情况与口腔扁平苔藓和口腔鳞状细胞癌患者临床特征的关系,采用Pearson相关分析法分析口腔扁平苔藓及口腔鳞状细胞癌患者中Galectin-9和TIM3表达的相关性。结果口腔鳞状细胞癌组织中Galectin-9的阳性表达率低于口腔扁平苔藓组织和正常口腔黏膜组织,TIM3的阳性表达率高于正常口腔黏膜组织,差异均有统计学意义(P﹤0.05);口腔扁平苔藓组织中Galectin-9的阳性表达率低于正常口腔黏膜组织,TIM3的阳性表达率高于正常口腔黏膜组织,差异均有统计学意义(P﹤0.05)。糜烂型口腔扁平苔藓患者口腔扁平苔藓组织中Galectin-9的阳性表达率低于非糜烂型口腔扁平苔藓患者,TIM3的阳性表达率高于非糜烂型口腔扁平苔藓患者,差异均有统计学意义(P﹤0.05)。不同分化程度、淋巴结转移情况口腔鳞状细胞癌患者口腔鳞状细胞癌组织中Galectin-9的表达情况比较,差异均有统计学意义(P﹤0.05);不同分化程度口腔鳞状细胞癌患者口腔鳞状细胞癌组织中TIM3的表达情况比较,差异有统计学意义(P﹤0.05)。Pearson相关分析结果显示,口腔扁平苔藓和口腔鳞状细胞癌患者中Galectin-9和TIM3的表达均呈负相关(r=-0.415、-0.437,P﹤0.01)。结论Galectin-9和TIM3可能参与口腔扁平苔藓及口腔鳞状细胞癌的发生发展,并可能与口腔扁平苔藓的癌变有关。     

Shh和Ptch在食管鳞状细胞癌中的表达研究

目的探讨食管鳞状细胞癌中Shh和Ptch的表达及其意义。方法应用免疫组织化学方法检测35例食管癌及癌旁组织中shh和Ptch的表达情况。结果食管鳞癌中Shh和Ptch的表达阳性率分别为74．3％和68．6％，与正常组织相比Shh和Ptch在食管鳞癌组织中高表达。结论Shh和Ptch的高表达及Hh通路的激活参与了食管鳞癌的发生，可能足食管癌治疗的一个理想靶标。     

音猬因子、Ptch1基因在胃癌中的表达及临床意义

目的:研究胃癌组织中音猬因子（Shh）、Patched1(Ptch1)基因的表达及意义。方法:采用反转录-聚合酶链反应(RT-PCR)和Westem blot检测98例胃癌组织和配对的癌旁组织中Shh、Ptch1 mRNA和蛋白的表达情况。结果:RT-PCR检测结果显示,Shh、Ptch1 mRNA在胃癌中相对表达量分别为0.687±0.057、0.594±0.046,在胃癌旁组织中分别为0.314±0.025、0.293±0.074（P＜0.05）。Western blot检测结果显示,Shh、Ptch1蛋白在胃癌中的相对表达量分别为0.525±0.057、0.481±0.046,在胃癌旁组织中分别为0.236±0.025、0.215±0.074。胃癌组织Shh、Ptch1 mRNA和蛋白平均表达水平高于癌旁组织,差异有统计学意义（P＜0.05）;Shh、Ptch1 mRNA和蛋白表达水平与胃癌的分化程度、TNM分期、浸润深度和淋巴结转移明显相关（P＜0.05）,与患者年龄、性别和肿瘤直径无明显相关（P＞0.05）。结论:胃癌组织中Shh、Ptch1蛋白呈高表达,Shh、Ptch1蛋白的高表达可能与胃癌的发生及发展有关。     

bcl-2蛋白在食管鳞状细胞癌及癌旁组织中的表达及其意义

目的 研究凋亡相关基因bcl-2在食管鳞状细胞癌及癌旁组织中的表达及其意义.方法 应用EnVision二步法检测62例食管鳞状细胞癌及癌旁组织中bcl-2蛋白的表达.结果 bcl-2在单纯增生、高级别上皮内瘤变及鳞状细胞癌组织中表达率分别为80.3 ％(49/61)、45.9 ％(28/61)及67.7％(20/62),而在正常食管黏膜组织中仅少量表达[3.3％(1/30)],正常黏膜组与其余各组间表达率差异有统计学意义(x2=54.437,P＜0.01).bcl-2蛋白表达与食管鳞状细胞癌的组织分化程度、浸润深度无相关性(x2=0.219、x2=5.878,均P＞0.05),但bcl-2蛋白的表达与淋巴结转移有相关性(x2=4.120,P＜0.05).结论 bcl-2可能在食管癌早期阶段发挥作用,其可能成为判断食管鳞状细胞癌预后的指标之一.     

口腔鳞癌组织中KAI1 mRNA的表达

目的:探讨肿瘤转移抑制基因KAI1 mRNA在口腔鳞状细胞癌中的表达及其临床病理意义。方法:采用原位杂交法检测74例原发性口腔鳞癌(无淋巴结转移49例,有淋巴结转移25例)、21例相应的淋巴结转移灶、19例口腔黏膜白斑和10例正常口腔黏膜石蜡标本组织中KAI1 mRNA的表达情况。结果:KAI1 mRNA在正常黏膜组、黏膜白斑组、鳞癌无淋巴结转移组、有淋巴结转移组和淋巴结转移灶组中阳性表达率分别为100%、84.2%、61.2%、28.0%和4.8%,其中无淋巴结转移鳞癌组显著低于正常和黏膜白斑组,P<0.05,淋巴结转移组显著低于无淋巴结转移组,P=0.045,淋巴结转移灶组显著低于相应鳞癌组,P=0.040。低分化鳞癌的KAI1阳性表达率显著低于高、中分化鳞癌,P<0.001。KAI1的表达与鳞癌患者的年龄、性别、肿瘤大小和浸润深度以及pT-MN分期无显著相关,P>0.05。结论:KAI1 mRNA表达下调可能与口腔鳞状细胞癌的组织分化程度和淋巴结转移有关。     

Bub1在食管鳞状细胞癌组织和正常食管黏膜的表达及临床意义

目的：观察Bub1 mRNA及Bub1蛋白在食管鳞状细胞癌和正常食管黏膜组织中的表达差别,探讨Bub1在食管鳞状细胞癌发生、发展中的作用及其临床意义。方法：收集40例食管鳞状细胞癌标本及相应的食管切缘正常黏膜组织,利用免疫组化染色及原位杂交技术检测Bub1蛋白和Bub1 mRNA在食管鳞状细胞癌组织和正常食管黏膜组织中的表达。结果：与正常黏膜组织相比较,Bub1 mRNA及Bub1蛋白在食管鳞状细胞癌组织中表达水平明显降低（P〈0.005）,且Bub1 mRNA及Bub1蛋白的表达水平与食管鳞状细胞癌分化程度及有无淋巴结转移相关（P〈0.05）。结论：Bub1表达水平高低可能与食管鳞状细胞癌的发生发展及淋巴结转移具有一定关系。     

音猬因子、Smo基因在宫颈鳞状细胞癌中的表达及临床意义

目的:探讨音猬因子(Shh)、Smoothened(Smo)基因在宫颈鳞状细胞癌中的表达及意义.方法:采用反转录-聚合酶链反应(RT-PCR)和Westem blot检测76例宫颈鳞状细胞癌组织和42例正常宫颈组织中Shh、Smo mRNA和蛋白的表达情况.结果:RT-PCR检测结果显示,Shh、Smo mRNA在宫颈鳞状细胞癌中相对表达量分别为0.715±0.048、0.638±0.037,在正常宫颈组织中分别为0.341±0.072、0.311±0.051(P<0.05).Western blot检测结果显示,Shh、Smo蛋白在宫颈鳞状细胞癌中的相对表达量分别为0.568±0.013、0.521±0.056,在正常宫颈组织中分别为0.281±0.047、0.252±0.064(P<0.05).宫颈鳞状细胞癌组织Shh、Smo mRNA和蛋白平均表达水平高于正常宫颈组织,差异有统计学意义(P<0.05);Shh、Smo mRNA和蛋白表达水平与宫颈鳞状细胞癌的分化程度明显相关(P<0.05),与患者年龄、FIGO分期、侵犯血管和神经、淋巴结转移、远处转移无明显相关(P>0.05).结论:宫颈鳞状细胞癌组织中Shh、Smo蛋白呈高表达,Shh、Smo蛋白的高表达可能与宫颈鳞状细胞癌的发生、发展及转移关系密切.     

Acquired mutations in the MXR/BCRP/ABCP gene alter substrate specificity in MXR/BCRP/ABCP-overexpressing cells

A disparity was noted in the transport of rhodamine 123 among nine MXR/BCRP/ABCP-overexpressing cells studied; all demonstrated mitoxantrone transport, whereas only two effluxed rhodamine 123. When the MXR/BCRP/ABCP gene was sequenced in the cell lines studied, differences were noted at amino acid 482, predicted to be at the start of the third transmembrane domain. Sequencing genomic DNA revealed wild-type MXR/BCRP/ABCP to have an arginine at position 482. Cells having a threonine or glycine at position 482 were able to efflux rhodamine 123, whereas cells having an arginine were not. A vaccinia virus expression system confirmed that rhodamine as well as doxorubicin efflux is observed with R482T or R482G but not with the wild-type R482; all three MXR/BCRP/ABCP forms transported mitoxantrone. Cross-resistance studies suggest that, compared with wild-type MXR/BCRP/ABCP, cells having an R482T mutation have higher anthracycline resistance, whereas an R482G mutation seems to confer relatively less resistance to SN-38 and topotecan. These results suggest that amino acid 482 has a crucial role in MXR/BCRP/ABCP function and that mutation of a single amino acid residue significantly changes substrate specificity, thus altering the drug resistance phenotype.     

MicroRNAs involved in the EGFR/PTEN/AKT pathway in gliomas

Gliomas are the most common type of malignant primary brain tumor. Despite advances in surgery, radiation therapy, and chemotherapy, the prognosis of patients with gliomas has not significantly improved. MicroRNAs (miRNAs), a class of non-coding RNAs, 21–25 nucleotides long, negatively regulate the expression of target genes by interacting with specific sites in mRNAs, and play a critical role in the development of gliomas. The EGFR/PTEN/AKT pathway is a promising target for anti-glioma therapy. Recent studies have showed that regulation of the EGFR/PTEN/AKT pathway by miRNAs plays a major role in glioma progression, indicating a novel way to investigate the tumorigenesis, diagnosis, and therapy of gliomas. Here, we focus on recent findings of miRNAs with respect to the EGFR/PTEN/AKT pathway in gliomas.     

Thalidomide/estramustine/paclitaxel in metastatic androgen-independent prostate cancer

Background

This is a phase I/II trial of thalidomide with estramustine and paclitaxel in men with androgen-independent prostate cancer (AIPC) who underwent previous chemotherapy.

Patients and Methods

Men with progressive AIPC were treated with oral thalidomide (200 mg, 400 mg, or 600 mg daily), intravenous paclitaxel (100 mg/m² over 3 hours on days 3 and 10), and oral estramustine (140 mg 3 times daily on days 1-5 and days 8-12) every 21 days.

Results

Phase I: first cycle dose-limiting toxicity occurred in 0 of 3 patients at 200 mg thalidomide daily, 0 of 3 at 400 mg daily, and 1 of 3 at 600 mg daily (the designated maximum tolerated dose). Phase II: twenty-nine of 38 evaluable patients (76%; 95% confidence interval, 67%-87%) experienced a 50% decrease in prostate-specific antigen level. Five of 18 patients (28%) with measurable disease exhibited an objective response. Nine of 14 patients (64%) with disease refractory to previous taxane therapy had 50% decreases in prostate-specific antigen level. Grade 3/4 adverse events included neutropenia (9 of 39 [23%]), fatigue (9 of 39 [23%]), dyspnea (8 of 39 [21%]), and thromboembolic events (7 of 39 [18%]). Cumulative dose-limiting toxicity rates were minimal (13%) with thalidomide at 200 mg daily.

Conclusion

The profile of activity of thalidomide/paclitaxel/estramustine in taxane-refractory AIPC warrants further investigation.     

Ras/Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR inhibitors: rationale and importance to inhibiting these pathways in human health

The Ras/Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR cascades are often activated by genetic alterations in upstream signaling molecules such as receptor tyrosine kinases (RTK). Integral components of these pathways, Ras, B-Raf, PI3K, and PTEN are also activated/inactivated by mutations. These pathways have profound effects on proliferative, apoptotic and differentiation pathways. Dysregulation of these pathways can contribute to chemotherapeutic drug resistance, proliferation of cancer initiating cells (CICs) and premature aging. This review will evaluate more recently described potential uses of MEK, PI3K, Akt and mTOR inhibitors in the proliferation of malignant cells, suppression of CICs, cellular senescence and prevention of aging. Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways play key roles in the regulation of normal and malignant cell growth. Inhibitors targeting these pathways have many potential uses from suppression of cancer, proliferative diseases as well as aging.