VEGF-C在晚期卵巢癌原发灶和转移灶中的表达比较

目的 研究血管内皮生长因子C（VEGV-C）在晚期卵巢癌原发灶和转移灶中的表达并进行比较,探讨其与晚期卵巢癌发生、发展及预后的关系。方法 应用SP免疫组织化学技术检测52例晚期卵巢癌原发灶以及相应的大网膜转移灶中VEGF-C的表达。结果 VEGF-C在52例卵巢上皮癌中原发灶的高表达率为21％（11／52）,其中透明细胞癌与黏液性癌相比有显著性差异（X^2=6．97,P〈0．01）;在转移灶的高表达阳性率为35％（18／52）,各种组织学类型之间无显著性差异（P〉0．01）。卵巢癌原发灶和转移灶之间VEGF-C表达无明显差异（P〉0．05）。结论 VEGF-C在晚期卵巢癌组织中表达增高,原发灶与转移灶之间无明显差异。     

CD138和乙酰化肝素酶在肝细胞癌中表达的组织芯片研究

目的探讨人肝细胞癌中CD138和乙酰化肝素酶的表达水平及其与肝细胞癌发生、发展、转移和复发的关系。方法运用组织芯片技术,通过免疫组织化学EnVision法研究197例肝细胞癌、癌旁肝组织及其中66例转移病灶中CD138和乙酰化肝素酶的表达水平。48例获得4～72个月随访。结果（1）CD138的表达率在肝细胞癌和癌旁肝组织中分别为48．7％（96／197）和65．0％（128／197。P〈0．05）,在早期和中晚期肝细胞癌中分别为61．7％（29／47）和44．7％（67／150,P〈0．05）,在有和无转移的中晚期肝细胞癌组中分别为33．3％（22／66）和53．6％（45／84,P〈0．05）,在1年之内和1年之上复发组中分别为23．3％（7／30）和61．1％（11／18,P〈0．05）;（2）乙酰化肝素酶的阳性表达率在肝细胞癌和癌旁肝组织中分别为35．5％（70／197）和12．7％（25／197,P〈0．05）,在早期和中晚期肝细胞癌中分别为29．8％（14／47）和37．3％（56／150,P〉0．05）,在伴随和不伴转移的肝细胞癌组织中48．5％（32／66）和28．6％（24／84,P〈0．05）,在1年之内和1年之上复发组中分别为50．0％（15／30）和44．4％（8／18,P〉0．05）;（3）66例伴随转移的肝细胞癌组织中,CD138在乙酰化肝素酶阴性组中的高表达率高于阳性组（P〈0．05）。结论（1）CD138在肝细胞癌组织中的低表达与其发生、发展、转移和复发密切相关,与分级、HBV感染及血清甲胎球蛋白值无关;（2）CD138蛋白低表达与乙酰化肝素酶高表达参与肝细胞癌的转移过程。     

窖蛋白-1在肺癌中的表达及意义

目的 探讨窖蛋白-1（caveolin-1）在不同类型肺癌组织中的表达及其与微血管密度（MVD）和临床病理因素之间的关系。方法 对154例原发性肺癌、相应癌旁正常肺组织及36例淋巴结转移癌行caveolin-1免疫组织化学染色;对154例原发性肺癌行CD34免疫组织化学（SP法）染色并进行微血管密度计数;Western印迹法检测其中50例新鲜肺癌组织及其癌旁正常肺组织中caveolin-1的表达情况。结果 caveolin-1为膜／质表达蛋白,在正常支气管上皮细胞和肺泡上皮细胞中的阳性率为100％。在肺癌组织中的阳性率为59．1％（91／154）,低于癌旁正常肺组织,P＜0．01;Western印迹结果进一步证实caveolin-1在肺鳞癌、肺腺癌组织中的表达均显著低于癌旁正常肺组织,P＜0．01。caveolin-1在小细胞肺癌（SCLC）和非小细胞肺癌（NSCLC）中的阳性率分别为7．1％和64．3％,二者间差异有统计学意义,P＜0．01。NSCLC中,有淋巴结转移组caveolin-1表达高于无淋巴结转移组（P=0．005）;Ⅲ、Ⅳ期组caveolin-1表达显著高于Ⅰ、Ⅱ期组（P=0．042）,caveolin-1表达与NSCLC的其他临床病理因素及MVD值无关（P＞0．05）。结论caveolin-1其作为一种肿瘤抑制因子的同时,可能还具有促进NSCLC进展和转移的活性。     

蛋白酶激活受体-1促进肺癌细胞侵袭和转移功能的研究

目的研究蛋白酶激活受体1（PAR-1）对人类肺癌细胞侵袭和转移功能的影响。方法采用阳离子脂质体介导法,将正义和反义PAR-1重组质粒“pC／PARls”和“pC／PARlas”分别转染至人肺巨细胞癌低转移株（PLA801C）和高转移株（PLA801D）细胞中;采用逆转录．聚合酶链反应（RT-PCR）和Western印迹检测转染后PAR-1基因和蛋白表达水平变化;通过MTT、软琼脂集落形成、流式细胞仪、细胞-基质黏附和Transwell细胞侵袭实验检测PAR-1对肺癌细胞转移相关功能的影响。结果转染正义和反义PAR-1分别明显上调和下调了PLA801C和PLA801D的PAR-1 mRNA和蛋白表达水平。转染正义PAR-1对细胞的生长和克隆形成具有促进作用;并可明显增强细胞对细胞外基质的黏附和侵袭能力（与空载体对照组比较均P〈0．01）。相反,转染反义PAR-1对细胞模型的生长和克隆形成具有抑制作用;能明显降低细胞的S期和G2／M期（与空载体对照组比较分别为P〈0．05、0．01）、升高G0／G1期所占比例（P〈0．01）;还可使细胞对细胞外基质的黏附力（P〈0．05）和侵袭力明显降低（P〈0．01）。结论正义和反义PAR-1基因能够分别上调和下调PAR-1的表达水平;PAR-1能够影响肺癌细胞的生长和增殖、黏附和侵袭特性。抑制PAR-1的表达可能是一种治疗肺癌的途径。     

胃腺癌组织中miR-23a与转移抑制因子1的表达及意义

目的：探讨miR-23a与转移抑制因子1（MTSS1）在胃腺癌组织中的表达及其临床意义。方法：运用荧光素报告载体系统检测miR-23a直接调控的靶基因并采用Transwell侵袭实验检测miR-23a对人胃腺癌细胞的侵袭能力。收集胃腺癌患者手术标本46例,癌旁组织作为正常对照,分别运用原位杂交,免疫组织化学EnVision法检测胃腺癌组织及癌旁组织中miR-23a,MTSS1的表达水平,并对二者进行相关性分析。结果：MTSS1是miR-23a直接调控的靶基因,miR-23a下调MTSS1蛋白的表达,增强胃癌细胞的侵袭能力。在46例胃腺癌组织中miR-23a阳性表达率为87．0％（40／46）,MTSS1蛋白阳性表达率为17．4％（8／46）,分别与癌旁对照组比,差异有统计学意义（P〈0．01）;miR-23a的表达随胃腺癌临床分期演化（P〈0．05）和浸润深度增加（P〈0．01）,且有淋巴结转移的miR-23a的表达显著高于无淋巴结转移组（P〈0．05）;MTSS1l的表达随胃腺癌临床分期演化（P〈0．05）和浸润深度增加而下调（P〈0．05）,且有淋巴结转移的MTSS1的表达显著低于无淋巴结转移组（P〈0．01）;相关性分析表明,miR-23a表达与MTSS1的表达呈显著负相关（r=-0．594,P〈0．05）。结论：miR-23a通过靶向抑制MTSS1促进胃腺癌细胞生长,侵袭转移;miR-23a的高表达和MTSS1蛋白低表达可能是胃腺上皮恶性转化以及胃癌发生浸润转移的重要生物学标志,检测二者对预测胃癌侵袭及转移有重要意义。     

Six1和Six4在食管鳞状细胞癌组织中的表达及其与预后的关系北大核心CSCD

目的探讨Six1和Six4在食管鳞状细胞癌（简称鳞癌）组织中的表达及其与临床病理特征及预后的关系。方法应用组织芯片技术、免疫组织化学EnVision法检测Six1和Six4在292例食管鳞癌组织及其相应的癌旁正常食管上皮组织中的表达水平,分析两者表达与临床病理特征及预后之间的关系。结果在292例食管鳞癌患者中,Six1、Six4蛋白在癌组织的阳性率分别为72．9％（213／292）和56．2％（164／292）,明显高于癌旁正常组织的阳性率,分别为33．2％（97／292）和32．5％（95／292）,差异具有统计学意义（P〈0．05）。X2检验结果显示Six1蛋白表达与肿瘤大小、肿瘤浸润深度及生存状态相关,其在死亡组患者的阳性率82．3％（130／158）,明显高于生存组阳性率61．9％（83／134,P〈0．05）。Six4蛋白表达与肿瘤分化程度及肿瘤浸润深度相关,肿瘤分化程度越高、浸润深度越深其阳性率越高。单因素Log-rank分析显示性别、TNM分期、Six1蛋白表达与食管鳞癌患者的总生存率相关（P〈0．05）。Six1蛋白表达阴性组5年生存率51．9％（41／79）高于阳性组的43．7％（93／213）,差异具有统计学意义（X^2=4．079,P=0．043）,其他临床参数与预后均未显示统计学意义（P〉0．05）。多因素Cox比例风险模型分析显示,TNM分期、Six1蛋白表达水平是影响食管癌术后患者预后的独立因素（P〈0．05）。结论Six1和Six4蛋白在食管鳞癌组织中均高表达,其表达水平与食管鳞癌发生发展及预后密切相关,Six1蛋白高表达是食管鳞癌患者预后不良的因素,可作为临床预测食管鳞癌患者预后的一个重要指标。     

Tiam-1 mRNA及其蛋白的表达与鼻咽癌浸润转移的关系

目的探讨Tiam-1 mRNA及其蛋白表达与鼻咽癌浸润转移的关系及意义。方法41例不同临床分期人鼻咽癌组织以及对照组20例鼻咽慢性炎症黏膜组织标本,用RT-PCR方法检测Tiam-1 mRNA表达水平,用免疫组化检测Tiam-1蛋白表达。结果Tiam-1 mRNA在鼻咽癌组织的平均表达水平为I．83±0．73,明显高于鼻咽黏膜慢性炎组织（0．87±0．45）（P〈0．01）;而且鼻咽癌组Tiam-1 mRNA高表达率为46．34％,明显高于对照组（P〈0．01）。Tiam-1蛋白在鼻咽癌组阳性率为65．85％（27／41）,明显高于对照组（15％,3／20）（P〈0．01）;其高表达率为43．90％（18／41）,明显高于对照组（P〈0．01）。Tiam-1 mRNA及其蛋白高表达率在临床TNM各分期间的差异均有明显统计学意义（P〈0．01）;在各T分期之间的差异亦有明显统计学意义（P〈0．01）。鼻咽癌中有淋巴结转移组Tiam-1 mRNA及其蛋白高表达率分别为59．26％（16／27）及55．56％（15／27）,均明显高于无淋巴结转移组的21．43％（3／14）及14．28（2／14）,（P〈0．05）。结论Tiam-1 mRNA及其蛋白高表达与鼻咽癌浸润转移有密切关系,提示Tiam-1基因可能是鼻咽癌的重要促浸润转移因子,有望成为治疗的靶标及有价值的预后判断指标。     

SEL1L基因在食管癌中的表达及意义

目的探讨SEL1L（human Sel-1-like）mRNA及其蛋白在食管癌组织中的表达及其临床意义。方法应用免疫组织化学SP法检测90例手术切除的食管鳞状细胞癌、35例距癌灶边缘5am以上切缘的正常黏膜、60例癌旁黏膜及20例内窥镜活检的食管鳞状上皮不典型增生组织中SEL1L蛋白的表达;运用原位分子杂交技术检测上述癌组织、正常黏膜、癌旁黏膜中SEL1L mRNA的表达。结果（1）SEL1L mRNA在食管鳞状细胞癌的表达率为80．0％（72／90）,较正常黏膜的14．3％（5／35）和癌旁黏膜的16．7％（10／60）高（P〈0．01）;SEL1L mRNA在有淋巴结转移组的表达阳性率为92．7％（38／41）比无淋巴结转移组69．4％（34／49）高（P〈0．01）。（2）SEL1L蛋白在鳞状细胞癌的表达阳性率为87．8％（79／90）,在鳞状上皮不典型增生中的表达阳性率为90．0％（18／20）。分别较正常黏膜的14．3％（5／35）和癌旁黏膜的13．3％（8／60）高（P〈0．01）。SEL1L蛋白表达与患者性别、年龄、肿瘤位置、大小、分化程度、浸润深度、淋巴结转移及临床分期均无明显相关性（P〉0．05）。（3）食管鳞状细胞癌组织中SEL1L mRNA和SEL1L蛋白的表达呈明显正相关（r=0．492,P〈0．01）。结论（1）SEL1L蛋白表达的调控主要在转录水平,SEL1L蛋白表达水平的升高主要是相应转录水平上调的结果。（2）SEL1L蛋白过表达可能是食管鳞状细胞癌发生的早期表现,SEL1L蛋白的检测可作为识别食管癌高风险患者的生物标记物。     

大肠癌及其转移灶中MMP-2和TIMP-2的表达和意义

目的研究基质金属蛋白酶-2（MMP-2）及其抑制物组织基质金属蛋白酶抑制剂-2（TIMP-2）在大肠癌进展中的作用及其临床意义。方法通过免疫组化的方法检测104例大肠癌组织及其转移灶中MMP-2和TIMP-2的表达。结果MMP-2在转移灶中的阳性率（72．1％）显著高于原发灶中的阳性率（53．8％）（P〈0．05）,而TIMP-2在转移灶中的阳性率（35．6％）显著低于原发灶中的阳性率（66．3％）（P〈0．05）;MMP-2和TIMP-2在大肠癌原发及转移灶中表达均具有相关性,且呈负相关。结论MMP-2和TIMP-2的表达与大肠癌转移密切相关,而且转移灶癌细胞MMP-2的表达明显增强,TIMP-2的表达明显下降,可作为临床判断大肠癌恶性程度、转移及预后的重要参考指标。     

膀胱移行细胞癌中fhit基因和survivin基因的表达和意义

目的探讨fhit基因和survivin基因在膀胱移行细胞癌中的表达和意义。方法用免疫组化S-P法检测62例膀胱移行细胞癌组织及10例正常膀胱粘膜组织中fhit蛋白和survivin蛋白的表达。结果10例正常膀胱粘膜组织中fhit蛋白表达均为阳性，survivin蛋白表达均为阴性；fhit蛋白在膀胱移行细胞癌中阳性表达率为46．77％（29／62），肿瘤不同分级中随恶性程度的增长表达减少，Ⅰ级与Ⅲ级比较，差异有统计学意义（P〈0．05），不同临床分期中随分期的增长表达减少，Tis～T1与T2～T4比较，差异无统计学意义（P〉0．05）；survivin蛋白在膀胱移行细胞癌中阳性表达率为56．5％（35／62），肿瘤不同分级中随恶性程度的增高表达增高（P〈0．05），不同临床分期中随分期的增长表达增高，差异有统计学意义（P〈0．05）；fhit蛋白和survivin蛋白表达相关（P〈0．05）。结论Fhit基因和survivin基因在膀胱移行细胞癌的发生、发展过程中起着重要的作用。Fhit基因可能通过肿瘤凋亡抑制途径发挥作用的。     

肺癌中凝血酶受体-1的表达及意义

目的 分析凝血酶受体(PAR-1)在肺癌组织及细胞中的表达情况，探讨PAR-1与人肺癌生物学行为的关系。方法 采用免疫组织化学、Westem blot及RT-PCR方法检测不同类型肺癌组织及细胞系中PAR-1的表达情况。结果 免疫组化S-P法检测PAR-1在52例肺癌组织中表达情况分别为：腺癌14／20例、鳞癌6／16例、大细胞癌6／10例、小细胞癌中4／6例。免疫组化阳性细胞多位于癌巢周边，1例侵及支气管软骨处的鳞癌癌巢及另1例低分化腺癌脉管内的癌栓呈明显阳性，1例腺癌组织周围的肺泡上皮不典型增生灶呈明显阳性。Westem blot及RT-PCR方法检测结果均显示PAR-1在人肺巨细胞癌细胞系高转移株(PLA801D)中的高表达明显高于其低转移株(PLA801C)。结论 PAR-1在各种组织类型的人肺癌组织中均有不同比例表达。PAR-1在人肺巨细胞癌细胞系高转移株(PLA801D)中的表达明显高于其低转移株(PLA801C)。PAR-1表达可能与肺癌的恶性表型及生物学行为有关。     

Expression of protease-activated receptors 1 and 2 in melanocytic nevi and malignant melanoma

Protease-activated receptors (PARs) are members of the G protein-coupled receptor superfamily that are activated by the proteolytic cleavage of their amino terminal domain. PAR-1 activation by thrombin results in several biologic effects, including platelet adhesion to other cells or extracellular matrix, fibroblast, and endothelial cell growth, whereas PAR-2, activated by trypsin, has mainly a proinflammmatory and angiogenetic role. PAR-1 and PAR-2 modulate cell proliferation in physiopathologic cell invasion processes, suggesting that they may play a role in the setting of cancer growth and metastasis. Here, we have investigated the expression of PAR-1 and PAR-2 proteins by immunohistochemistry in a series of benign and malignant melanocytic lesions: 20 melanocytic lesions (10 common melanocytic nevi and 10 atypical or "dysplastic" melanocytic nevi) and 50 melanomas (10 in situ melanomas, 10 melanomas T1, 10 melanomas T2, 10 melanomas T3 to T4, and 10 metastatic melanomas). PAR-1 was significantly overexpressed in atypical nevi and melanomas in comparison with common melanocytic nevi. PAR-2 was strongly and diffusely expressed by immunohistochemistry in all melanocytic lesions, with no statistically significant differences between nevi and melanomas. Because we found a differential expression in PAR-1 protein, but not in PAR-2, we next investigated the expression of PAR-1 messenger RNA (mRNA) by ribonuclease protection assay in paraffin-embedded tissues using a paraffin block RNA isolation procedure. Similarly to immunohistochemical results, PAR-1 mRNA expression was significantly higher in atypical nevi and melanomas in comparison with common nevi and controls. Overexpression of PAR-1 in atypical nevi and melanomas supports a role for PAR-1 in the initial phases of melanoma development as well as in tumor progression and metastasis. Conversely, the significance of PAR-2 up-regulation in both benign and malignant melanocytic lesions requires further research.     

NM-23 gene loss of heterozygosity and protein expression in high-stage laryngeal squamous cell carcinomas.

Tumor suppressor genes that reduce metastatic potential have been described in a variety of different tumor types. One of the main tumor metastasis suppressor genes is nm-23, which is a nucleoside diphosphate kinase. Two isotypes, nm-23H1 and nm-23H2, have been cloned and map to chromosome 17q21.3. In a variety of tumors, including colon cancer and breast cancer, loss of expression of nm-23 is associated with lymph node metastasis. In other organ systems, however, this relationship is not seen. In head and neck squamous cell carcinomas (HNSCC), there have been conflicting results regarding the association between nm-23 protein expression and metastatic potential. To further explore the tumor metastasis suppressor function of nm-23 in HNSCC, we studied high-stage laryngeal carcinomas, tumors with and without cervical lymph node metastasis for nm-23 protein expression and loss of heterozygosity of the gene locus. Twenty-five cases were included (11 cases with and 14 cases without metastasis). Loss of heterozygosity for the nm-23 gene locus was seen in 7 of 22 (32%) informative tumors. Using immunohistochemistry, most tumors expressed nm-23, though decreased expression was seen in 10 of 25 (40%) cases. Only 2 tumors showed negative expression. We did not find a correlation between either protein expression or loss of heterozygosity with metastatic disease or any other adverse prognostic factors in this group of high-stage laryngeal squamous cell carcinomas. These data imply that nm-23 may be tumor suppressor gene involved in HNSCC but that it may not function as a tumor metastasis suppressor in high-stage laryngeal carcinoma.     

Protease-activated receptor 2 signaling upregulates angiogenic growth factors in renal cell carcinoma

Renal cell carcinoma (RCC) is a highly vascular tumor associated with expression of various angiogenic growth factors. The precise process of how these growth factors are regulated in RCC is not fully understood. Recent evidence suggests that protease activated receptors (PARs), a new family of G-protein coupled receptors, play a crucial role in vascular development and tumor progression through a variety of mechanisms. However, the nature of PAR expression in human RCC tissues and its function in regulating angiogenesis in RCC are largely unknown. In this study, we investigated the expression and function of PAR-2 in RCC. RT-PCR and immunohistochemistry assays show that PAR-2 expression is significantly increased in human RCC tissue compared with the adjacent non-neoplastic kidney tissue. In RCC derived cells, PAR-2 is functional as evidenced by robust signaling through MAP kinases including ERK1/2 and JNK. Furthermore, activation of PAR-2 significantly upregulates several angiogenic cytokines, including interleukin-6 (IL-6), IL-8, monocytes chemotactic protein-1 (MCP-1) and growth-related oncogene (GRO). To our knowledge, this is the first report that characterized PAR-2 expression in RCC tissue and further demonstrated that PAR-2 has a critical role in regulating angiogenesis in RCC.     

Alpha-methylacyl-CoA racemase/P504S overexpression in colorectal carcinoma is correlated with tumor differentiation.

Alpha-methylacyl-CoA racemase (AMACR)/P504S is an enzyme involved in the metabolism of branched-chain fatty acids. Little is known about correlation of AMACR expression with colorectal carcinoma (CRC) differentiation and prognosis. We investigated the expression of AMACR in 106 cases of primary CRC, and in 47 lymph nodes with metastatic CRC by immunohistochemical analysis. These cases were divided into 3 groups according to the histologic differentiation of the primary tumors. group A included 50 cases of histologically well and moderately differentiated CRCs, 20 of these with lymph node metastasis; group B included 23 cases of well and moderately differentiated CRCs, histologically similar to group A, except these tumors had small foci (less than 20%) of high-grade carcinoma, and 10 of these had lymph node metastasis; group C included 33 cases of poorly differentiated adenocarcinoma and undifferentiated carcinoma, 17 with lymph node metastasis. The results showed the overall positive rates of expression in primary and metastatic CRCs were 59.4% and 46.8%, respectively. Expression in groups A (76.0%) and B (69.6%) was much higher than that in group C (27.3%). In group B, although overexpression of AMACR in primary tumors was similar to that of group A, it was only seen in 30.0% of group B metastatic tumors, which was similar to the rate of expression in group C (23.5%). In contrast, rates of expression in group A primary and metastatic tumors were similar (80.0% and 75.0%). Positive staining for AMACR in benign epithelium adjacent to tumor was rare (<2%). No relation was found between AMACR expression and overall survival. Our findings support the view that the expression of AMACR in CRC is correlated with tumor differentiation.     

结直肠癌nm23基因表达、突变与侵袭、转移的相关性

目的　探讨nm2 3基因异常表达及突变与结直肠癌侵袭、转移之间的关系。方法　应用免疫组化LDP法、流式细胞术、PCR SSCP法对 1 5 0例结直肠癌标本及正常黏膜组织和 1 6例大肠腺瘤分 3组进行nm2 3蛋白检测和基因突变的筛查。结果　①免疫组化发现 73例结直肠癌中nm2 3 H1蛋白表达水平低于正常黏膜组织和良性腺瘤 (P <0 0 0 1 ,P <0 0 0 1 ) ,转移癌表达低于原发癌 (P <0 0 0 1 ) ,结直肠癌nm2 3 H1蛋白表达与组织分化、肠壁浸润、Dukes分期、转移均密切相关 (P <0 0 0 1 ) ,低表达组术后预后差于高表达组 (P <0 0 0 1 ) ;②流式细胞术研究表明 ,nm2 3 H1蛋白在癌组织表达低于癌旁组织 ;在结直肠癌伴有转移 (3/ 1 4 )时其表达低于未发生转移者 (1 1 / 1 4 ) ;③nm2 3 H1基因第 2～ 5位外显子在 6 3例结直肠癌组织中未发现异常突变。结论　结直肠癌中nm2 3基因突变可能是小概率事件 ,在结直肠癌的侵袭、转移中不发挥主要作用 ,而nm2 3 H1蛋白表达降低则与结直肠癌的侵袭、转移有关 ,提示检测nm2 3蛋白表达是判断结直肠癌预后有价值的指标之一。     

miR-23a与转移抑制因子1在结肠癌中的表达及其临床意义

目的 探讨miR-23a( Homo sapiens miR-23a)与转移抑制因子1(metastasis suppressor 1,MTSS1)在结肠癌中的表达情况及其临床意义.方法 运用荧光素报告载体系统检测miR-23a直接调控的靶基因并采用Transwell侵袭实验检测miR-23a对人结肠癌SW620细胞的侵袭能力.收集结肠癌患者手术标本92例,癌旁组织作为正常对照,分别运用原位杂交、免疫组织化学EliVision法检测结肠癌组织及癌旁组织中miR-23a、MTSS1的表达水平,并对二者进行相关性分析.结果 MTSS1是miR-23a直接调控的靶基因,miR-23a下调MTSS1蛋白的表达,增强结肠癌细胞侵袭能力.在92例结肠癌组织中miR-23a阳性表达率为87.0％( 80/92),MTSS1蛋白阳性表达率为17.4％(16/92),分别与癌旁对照组比,差异有统计学意义(P ＜0.01);miR-23a的表达随着结肠癌临床分期演进(P =0.029)和浸润深度增加而增加(P=0.000),且有淋巴结转移的miR-23a的表达显著高于无淋巴结转移组(P=0.041);MTSS1的表达随着结肠癌临床分期演进(P=0.027)和浸润深度增加而下调(P=0.017),且有淋巴结转移的MTSS1的表达显著低于无淋巴结转移组(P=0.009);相关性分析表明,miR-23a表达与MTSS1的表达呈显著负相关(r=-0.594,P=0.013).结论 miR-23a通过靶向抑制MTSS1促进结肠癌SW620细胞生长、侵袭转移;miR-23a的高表达和MTSS1蛋白低表达可能是肠黏膜恶性转变以及结肠癌发生浸润转移的重要生物学标志,检测二者对预测结肠癌浸润转移有重要意义.     

Differential expression of protease-activated receptors-1 and -2 in stromal fibroblasts of normal, benign, and malignant human tissues

The serine proteases thrombin and trypsin are among many factors that malignant cells secrete into the extracellular space to mediate metastatic processes such as cellular invasion, extracellular matrix degradation, angiogenesis, and tissue remodeling. The degree of protease secretion from malignant cells has been correlated to their metastatic potential. Protease activated receptors (PAR)-1 and -2, which are activated by thrombin and trypsin respectively, have not been extensively characterized in human tumors in situ. We investigated the presence of PAR-1 and PAR-2 in human normal, benign and malignant tissues using immunohistochemistry and in situ hybridization. Our results demonstrate PAR-1 and PAR-2 expression in the tumor cells, mast cells, macrophages, endothelial cells, and vascular smooth muscle cells of the metastatic tumor microenvironment. Most notably, an up-regulation of PAR-1 and PAR-2 observed in proliferating, smooth muscle actin (SMA)-positive stromal fibroblasts surrounding the carcinoma cells was not observed in normal or benign conditions. Furthermore, in vitro studies using proliferating, SMA-positive, human dermal fibroblasts, and scrape-wounded human dermal fibroblasts demonstrated the presence of PAR-1 and PAR-2 not detected in quiescent, SMA-negative cultures. PAR-1 and PAR-2 in the cells forming the tumor microenvironment suggest that these receptors mediate the signaling of secreted thrombin and trypsin in the processes of cellular metastasis.