口腔鳞状细胞癌组织中VASH-1、LGR5的表达及相关性分析

目的分析口腔鳞状细胞癌组织中血管生成抑制蛋白-1(VASH-1)、富含亮氨酸重复序列G-蛋白偶联受体5(LGR5)的表达及临床意义。方法采用免疫组织化学SP法对2016年3月至2018年3月唐山市协和医院收治的87例口腔鳞状细胞癌患者癌组织及癌旁组织中VASH-1、LGR5的表达情况进行检测,分析VASH-1、LGR5表达与口腔鳞状细胞癌患者临床病理特征的关系,并分析口腔鳞状细胞癌组织中VASH-1、LGR5表达的相关性。结果口腔鳞状细胞癌组织中VASH-1阳性率、LGR5阳性率均显著高于癌旁组织(P 0. 05)。病理分级为Ⅰ级、肿瘤直径小于等于2 cm、TNM分期为Ⅰ～Ⅱ期、无淋巴结转移的口腔鳞状细胞癌患者VASH-1阳性率、LGR5阳性率均显著低于病理分级为Ⅱ～Ⅲ级、肿瘤直径大于2 cm、TNM分期为Ⅲ～Ⅳ期、淋巴结转移患者(P 0. 05)。经Spearman相关性分析显示:口腔鳞状细胞癌组织中VASH-1表达和LGR5表达呈正相关(Rs=0. 723,P=0. 000)。结论口腔鳞状细胞癌组织中VASH-1、LGR5呈明显高表达,其水平与肿瘤直径、病理分级、淋巴结转移存在密切相关性,且VASH-1表达和LGR5表达呈正相关。     

胰腺导管腺癌中DR5蛋白及mRNA的表达及临床意义

目的检测死亡受体5(death receptor 5,DR5)蛋白及mRNA在胰腺导管腺癌癌组织及癌旁正常组织中的表达及临床意义。方法采用免疫组化法检测DR5在117例胰腺导管腺癌石蜡包埋组织和癌旁正常组织中的表达,并对DR5蛋白表达与胰腺导管腺癌临床病理特征的关系进行系统性分析。同时利用qRT-PCR及Western blot法各检测20例配对的新鲜冻存的胰腺导管腺癌癌组织和癌旁正常组织中DR5蛋白及mRNA的表达。结果免疫组化结果显示,DR5主要表达于癌旁组织细胞的细胞膜中,DR5在胰腺导管腺癌组织中的阳性率为41. 02%(48/117),癌旁正常组织中的阳性率为74. 36%(87/117),两组表达差异有统计学意义(P 0. 001); DR5表达与分化程度(P 0. 05)、TNM分期(P 0. 05)、神经浸润(P 0. 001)以及淋巴结转移情况(P 0. 05)均有明显相关性。qRT-PCR及Western blot结果显示,癌旁正常组织中DR5蛋白及mRNA的表达量均远高于胰腺导管腺癌组织,两组相比差异有统计学意义(P 0. 001)。结论 DR5蛋白表达与胰腺导管腺癌分化程度、TNM分期、神经浸润及淋巴结转移均相关,对其进行检测可用于评估胰腺导管腺癌患者的病情严重程度及预后。     

PDCD4在喉癌中的表达及意义

目的研究PDCD4在喉癌组织中的表达及意义。方法应用免疫组化方法检测PDCD4蛋白在喉癌及癌旁组织中的表达。应用Real-time PCR方法检测PDCD4 mRNA在30例喉癌组织及癌旁正常组织中的表达,并与临床资料进行统计学分析。结果与癌旁组织相比,PDCD4蛋白在喉癌组织中表达水平明显降低,PDCD4 mRNA在喉癌组织中表达水平明显下调(0.05,t=4.714),与分期、分化及淋巴结转移明显相关,但与年龄、性别及分型不相关。结论 PDCD4在喉癌组织低表达,与分期、分化及淋巴结转移相关。     

口腔鳞癌组织中PTPRK基因的异常表达及甲基化分析

目的分析口腔鳞癌(oral squamous cell carcinoma,OSCC)中受体型蛋白酪氨酸磷酸酶K(protein tyrosine phosphatase,receptor type K,PTPRK)基因的甲基化状态及其mRNA和蛋白表达,探讨OSCC的发生、发展机制。方法收集OSCC组织57例及癌旁正常口腔黏膜组织41例,采用甲基化特异性PCR(methylation specific PCR,MSP)、RT-qPCR和免疫荧光染色分析OSCC组织及癌旁正常口腔黏膜组织中PTPRK基因甲基化状态、mRNA和蛋白的表达以及甲基化与临床病理特征之间的关系。结果 OSCC组织中PTPRK基因甲基化阳性率高于癌旁正常口腔黏膜(59. 65%vs 34. 15%,P 0. 05)。OSCC组织中PTPRK的mRNA和蛋白表达量明显低于癌旁正常口腔黏膜(mRNA水平:0. 53±0. 23 vs 1. 05±0. 08,P 0. 001;蛋白水平:0. 43±0. 21 vs 1. 02±0. 78,P 0. 01),OSCC组织中PTPRK基因mRNA的表达与PTPRK基因的甲基化状态相关(P 0. 01)。PTPRK基因的高甲基化状态与淋巴结转移、TNM分期及病理分级相关(P 0. 05)。结论 PTPRK基因启动子区Cp G岛高甲基化可能与OSCC的发生、进展相关,有可能成为OSCC早期诊断及药物治疗的分子靶点。     

食管鳞癌中Versican、NF-κB和MMP-9的表达及其与临床病理特征的关系

目的:观察食管鳞癌组织中多功能蛋白聚糖(Versican)、核因子-κB(Nuclear factor-kappa B,NF-κB)和基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)蛋白及 mRNA 的表达及其与临床病理特征的关系.方法:采用免疫组化及原位杂交方法检测 47 例食管鳞癌组织及癌旁正常食管黏膜组织中 Versican、NF-κB 和 MMP-9 蛋白及 mRNA 的表达情况,并分析其与临床病理特征的关系及三者的相关性.结果:食管鳞癌组织中 Versican、NF-κB 和 MMP-9 蛋白及 mRNA 表达的阳性率均明显高于癌旁正常食管黏膜组织中的表达(P＜0.05);食管鳞癌组织中 Versican、NF-κB 和 MMP-9 蛋白及 mRNA 的高表达与浸润深度越深、淋巴结有转移、TNM 分期越晚均有关(P＜0.05),与性别、年龄、组织分级均无关;食管鳞癌组织中,Versican、NF-κB 和 MMP-9 三者在蛋白及 mRNA 水平的表达均呈正相关(P＜0.05).结论:Versican、NF-KB 和 MMP-9 在食管鳞癌中的高表达与肿瘤的生成、浸润及转移密切相关,并且三者具有相关性,为我们进一步探索食管鳞癌浸润转移的潜在机制奠定了基础.     

结直肠癌中p-AKT和PTEN蛋白的表达及其临床意义

目的探讨磷酸化AKT(p-AKT)和PTEN蛋白在结直肠癌中的表达及其临床意义。方法应用免疫组化PV-9000两步法检测80例结直肠癌手术切除标本和20例癌旁正常组织中p-AKT、PTEN蛋白的表达,分析两者与临床病理特征的关系。结果 p-AKT蛋白在结直肠癌中表达的阳性率(71.25%)显著高于癌旁正常组织(10.00%,P0.05);而PTEN蛋白在结直肠癌中表达的阳性率(45.00%)与癌旁正常组织(100.00%,P0.05)相比具有统计学意义的降低,二者的表达与肿瘤的分化程度、TNM分期、浆膜浸润、淋巴结转移具有明显相关性(P0.05),并且p-AKT与PTEN蛋白的表达呈显著负相关(r=-0.314,P=0.005)。结论 p-AKT和PTEN蛋白的表达可能与结直肠癌的发生、发展密切相关。联合检测二者的表达,对判断结直肠癌的恶性程度及预后具有重要的临床意义。     

iASPP和caspase-9在食管癌组织中的表达及临床意义

目的探讨p53凋亡刺激蛋白家族抑制成员(iASPP)和caspase-9在食管癌中表达与食管癌临床病理特征的关系。方法收集临床食管癌标本85例及30例癌旁正常组织,采用免疫组织化学SP染色法和反转录PCR(RT-PCR)法检测iASPP和caspase-9在食管癌组织和癌旁正常组织中蛋白和mRNA表达情况,分析iASPP和caspase-9表达与食管癌临床病理特征之间的相关性。结果 SP法染色显示iASPP在食管癌组织和正常组织中的阳性表达率分别为72.9%(62/85)、16.7%(5/30),caspase-9在食管癌组织和正常组织中的阳性表达率分别为18.8%(16/85)、76.7%(23/30),二者差异均具有统计学意义(P0.01)。RT-PCR显示在食管癌中iASPP高表达和caspase-9低表达与食管癌的浸润深度、淋巴结转移、临床TNM分期相关(P0.05),而与食管癌患者的性别、年龄、肿瘤大小、组织类型均无相关性(P0.05),在食管癌组织中,iASPP和caspase-9的表达呈负相关(P0.05)。结论 iASPP和caspase-9在食管癌中的不同表达与食管癌的浸润深度、淋巴结转移、临床TNM分期显著相关,且二者呈负相关。     

哈族食管鳞癌中PLCE1基因启动子甲基化水平的实验研究

目的探讨PLCE1基因启动子甲基化与食管鳞癌发生发展的关系。方法分别用免疫组化(IHC)和甲基化特异PCR(MSP)方法检测51例新疆哈萨克族食管鳞癌及相应癌旁正常组织中PLCE1蛋白表达水平及其启动子甲基化水平,分析其与病理资料相关性;分别用Western blot及MSP检测食管鳞癌细胞在5-aza-dC作用前后PLCE1蛋白表达及启动子甲基化变化情况。结果新疆哈萨克族食管鳞癌组织中PLCE1蛋白表达高于癌旁正常组织,而其基因甲基化水平则较低(P 0.001),且蛋白高表达的癌组织中其甲基化水平低于蛋白低表达的癌组织(P=0.028),PLCE1甲基化水平与其蛋白表达水平具有明显的相关性(χ~2=4.791,P=0.028),并与患者的淋巴结及远处转移(χ~2=7.242,P=0.027)和TNM分期(χ~2=7.883,P=0.019)明显相关;在食管鳞癌细胞系中PLCE1甲基化程度同样与蛋白表达水平成反比,5-aza-dC处理可抑制TE-1和Kyse150的PLCE1甲基化,提高其蛋白表达。结论食管鳞癌组织中PLCE1甲基化低与其蛋白表达高、淋巴结和远处转移及TNM分期相关,5-aza-dC处理可抑制PLCE1甲基化程度,增高其蛋白表达。     

Sonic hedgehog mRNA及其蛋白在大肠癌的表达及其临床意义

目的 探讨大肠癌中Hedgehog信号通路相关基因Sonic hedgehog(SHH)的表达及其临床意义.方法 应用RT-PCR及免疫组织化学法(Envison二步法)检测本院2008年12月至2009年4月收集的43例大肠癌组织和距离癌10 cm以上的癌旁组织的SHHmRNA及蛋白的表达情况,并与20例非肠癌患者的正常大肠组织对照,分析其与大肠癌患者临床和病理各变量的相关性.结果 凝胶成像分析显示阳性表达的样本SHH mRNA片段大小为280 bp,与理论值相符,未表达的样本则未见相应条带.大肠癌组织SHHmRNA表达阳性率为27.9%(12/43),与癌旁组织的18.6%(8/43)差异无统计学意义(P＞0.05),但均显著高于正常大肠黏膜的0%(0/20)(均P＜0.05).SHH mRNA在大肠癌组织的表达强度显著高于癌旁组织(P＜0.05),在正常大肠黏膜则未见其表达.免疫组织化学显示SHH蛋白在大肠癌组织和癌旁组织均有阳性表达,细胞膜及胞质内出现棕黄褐色颗粒,背景不着色;正常大肠黏膜阴性表达.SHH蛋白在大肠癌组织的表达阳性率显著高于正常大肠黏膜[25.6%(11/43)比0%(0/20),P＜0.05],在大肠癌组织与癌旁组织、癌旁组织与正常大肠黏膜的表达阳性率差异则无统计学意义(均P＞0.05).SHH蛋白的表达强度大肠癌组织＞癌旁组织＞正常大肠黏膜(均P＜0.05).大肠癌组织中的SHH mRNA和蛋白表达强度与患者的年龄、性别、临床分期、肿瘤部位、肿瘤浸润深度、病理分型等变量均无明显关系(均P＞0.05).结论 SHH mRNA和蛋白在大肠癌组织中表达显著增强,但与临床和病理各变量无关联.     

Gab2在结直肠癌组织中的表达及其意义

目的:探讨Gab2在结直肠癌组织中的表达及其意义。方法:应用免疫组化方法检测78例结直肠癌及46例癌旁正常组织中Gab2的表达水平,并结合临床病理学资料进行统计学分析,Western blot方法检测10对结直肠癌组织及相对应的癌旁正常组织中Gab2的表达状况。结果:免疫组化检测结果显示78例结直肠癌组织中Gab2蛋白表达阳性率为53.85%,而癌旁正常组织中Gab2蛋白多不表达或弱表达,两组结果比较差异有显著性(P0.001);Gab2蛋白在结直肠癌中的表达与肿瘤TNM分期及有无淋巴结转移呈正相关(P0.05),与肿瘤大小、分化程度无明显相关性(P0.05);Western blot检测结果显示结直肠癌组织中Gab2蛋白表达显著高于相对应的癌旁正常组织(P0.05)。结论:Gab2在结直肠癌组织中过表达,可能在结直肠癌的发生发展中起重要作用。     

Genome-wide profiling of oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is a common malignancy, the incidence of which is particularly high in some Asian countries due to the geographically linked areca quid (AQ) chewing habit. In this study, array-based comparative genomic hybridization was used to screen microdissected OSCCs for genome-wide alterations. The highest frequencies of gene gain were detected for TP63, Serpine1, FGF4/FGF3, c-Myc and DMD. The highest frequencies of deletion were detected for Caspase8 and MTAP. Gained genes, classified by hierarchical clustering, were mainly on 17q21-tel; 20q; 11q13; 3q27-29 and the X chromosome. Among these, gains of EGFR at 7p, FGF4/FGF3, CCND1 and EMS1 at 11q13, and AIB1 at 20q were significantly associated with lymph node metastasis. The genomic profiles of FHIT and EXT1 in AQ-associated and non-AQ-associated OSCCs exhibited the most prominent differences. RT-PCR confirmed the significant increase of TP63 and Serpine1 mRNA expression in OSCC relative to non-malignant matched tissue. A significant increase in Serpine1 immunoreactivity was observed from non-malignant matched tissue to OSCC. However, there was no correlation between the frequent genomic loss of Caspase 8 and a significant decrease in Caspase8 expression. These data demonstrate that genomic profiling can be useful in analysing pathogenetic events involved in the genesis or progression of OSCC.     

Adenoid squamous cell carcinoma of the oral cavity

Because immunohistochemical features of adenoid squamous cell carcinoma (AdSCC) of the oral cavity is unclear, the author reports herein AdSCC in the gingival with an emphasis on immunohistochemical features. A 73-year-old woman presented with a left lower gingival tumor. The tumor was mildly elevated tumor measuring 1.5 x 1.5 x 0.5 cm. Dentist's diagnosis was granulation tissue, and a biopsy was taken. The biopsy showed proliferation of carcinoma cells arranged in cords, and squamous and tubular differentiations were noted in places. The biopsy diagnosis was adenosquamous carcinoma. Tumor excision with resection of mandibular bone was performed. The resected tissue showed a mixture and squamous cell carcinoma and tubular formation. Gradual merges between the two and acantholytic features of the squamous cell carcinoma element were seen. Both components were free from mucins. Both components were positive for pancytokeratins (AE1/3, CAM5.2) +++, cytokeratin (CK) 5/6 +, CK34βE12 ++, CK7 +, CK14 +++, CEA +, CA19-9 +, CA125 +, p53 +++, p63 +++, KIT + and MUC1 ++. Both components were negative for CK8, CK18, CK19, CK20, EMA, vimentin, TTF-1, desmin, myoglobin, S100 protein, melanosome, smooth muscle actin, CD34, CDX2, CD10, chromogranin, synaptophysin, NSE, CD56, lysozyme, CD68, MDM2, PDGFRA, MUC2, MUC5AC, and MUC6. Since both components were positive for squmaous cell carcinoma markers (CD5/6, CK34βE12, and p63) and adenocarcinoma markers (CEA, CA19-9, CA125, MUC1), this case of AdSCC appears an intermediate form between adenocarcinoma and squamous cell carcinoma. The margins were negative. No metastasis was found by imaging techniques. The patient is now free from tumor and is followed up carefully.     

Spindle cell squamous carcinoma of the oral region

Summary Six cases of spindle cell squamous carcinoma (SCSC) of the oral cavity were studied clinicopathologically, immunohistochemically and ultrastructurally to summarize the clinicopathological features of this rare neoplasm and to discuss the debatable histogenesis of the sarcomatoid component and the differential diagnosis of SCSC. The mean age of the patients was 72 years and the female to male ratio was 1:2. Four of them had a history of irradiation for pre-existing squamous cell carcinoma. One patient died of SCSC. While clinical and histological prognostic factors of SCSC could not be determined, it was shown that radical surgery resulted in good prognosis. The epithelial nature of the sarcomatoid component of SCSC was clearly revealed by a combination of immunohistochemical staining for keratins and electron microscopic demonstration of tonofilament-like filaments and/or desmosome-like structures. Together with electron microscopic evaluation of the tumour cells, immunohistochemical characterization of tumour cells using antibodies to keratin, vimentin, glial fibrillary acidic protein and S-100 protein is very helpful in differentiating SCSC from true spindle cell sarcoma, melanoma and malignant myoepithelioma. In the immunohistochemical differential diagnosis of SCSC, it is important to remember that SCSC should not be ruled out of the differential diagnosis by a positive reaction for vimentin in sarcomatoid tumour cells. Absence of staining for keratin in the sarcomatoid tumour cells does not always exclude SCSC, because some SCSCs show immunoreactivity of keratin in their sarcomatoid components only with some anti-keratin antibodies. Different kinds of anti-keratin antibodies should be applied in the differential diagnosis of SCSC.     

表皮生长因子受体在口腔鳞癌组织中的表达及意义

目的：研究表皮生长因子受体（ＥＧＦＲ）在口腔鳞癌组织中的表达及其临床病理学意义。方法：采用免疫组化Ｓ－Ｐ法对６５例不同分化的口腔鳞癌组织及１０例正常口腔粘膜组织进行ＥＧＦＲ检测。结果：ＥＧＦＲ在鳞癌组织中呈异质性表达,其阳性率（６１．５％）明显高于正常口腔粘膜组织（Ｐ〈０．０１）;ＥＧＦＲ表达状况与鳞癌组织学分级,区域淋巴结转移及患者预后间存在相关关系（Ｐ〈０．０１或０．０５）。结论：ＥＧＦＲ在口     

Expression of E-cadherin and vimentin in oral squamous cell carcinoma

The aim of the study is to determine the levels of E-cadherin, vimentin expression in tumor tissues from patients with oral squamous cell carcinoma (OSCC), and the relationship between the expression of E-cadherin, vimentin and epithelial-mesenchymal transition, in order to explore its values for predicting the invasion and metastasis of oral squamous cell carcinoma, short survival of patients in many types of cancer. E-cadherin and vimentin expression of 10 benign and 42 OSCC tumor tissues was examined by immunohistochemical staining. E-cadherin is positively expressed in normal oral mucosa epithelium, but vimentin expression is not found in normal oral mucosa epithelia; the E-cadherin and vimentin were expressed in 26 of 42 (61.9%) and 16 of 42 (38.1%), respectively. No statistically difference was found for E-cadherin and vimentin expression in patients with different age, gender and tumor location, E-cadherin and vimentin expression was significantly associated with lymph node metastasis and tissue location (P < 0.05); E-cadherin expression was also significantly associated with tumor stage (P < 0.05); there are significantly difference between infiltrative margin and central area in patients with oral squamous cell carcinoma for E-cadherin and vimentin positive expression (P < 0.05). E-cadherin and vimentin positive expression was associated with tumor metastasis of oral squamous cell carcinoma. Our study preliminarily confirmed that EMT phenomenon is existed during the development of oral squamous cell carcinoma. Co-evaluation of E-cadherin and vimentin might be a valuable tool for predicting OSCC patient outcome.     

Primary adenoid squamous cell carcinoma of the oral cavity

Adenoid squamous cell carcinoma (ASCC) is an uncommon but well-recognized variant of squamous cell carcinoma that was first described by Lever in 1947. ASCC has been reported to originate in the sun-exposed skin of the head and neck and in other sites. An additional case of ASCC is reported here. The patient was a 64-year-old Japanese woman who requested examination of a reddish lesion on the left floor of the mouth. The biopsy material was diagnosed as squamous cell carcinoma. Clinical examination showed a well-circumscribed, 20 x 10 mm-sized lesion, which was categorized as cT2cN0cm 0. Tumor resection was therefore performed. Histologically, most parts of the lesion were conventional squamous cell carcinoma in situ, but the invasive part consisted of ASCC with gland-like or reticular appearance. The latter part was negative for mucin staining. Immunohistochemically, this lesion was positive for pancytokeratin, high-molecular-weight keratin, cytokeratin (CK) 7/8, CK19, E-cadherin and p53, but negative for vimentin, CK20, and S-100 protein. The Ki-67 labeling index was 50.3% in the ASCC part and 34.5% in the carcinoma in situ part. These findings and a review of the literature indicate that a gland-like feature of ASCC is associated with the loss of cell adhesion in the center of the cancer nests, and it can be confirmed simply by mucin staining to be neither an adenosquamous carcinoma nor ductal involvement of conventional squamous cell carcinoma.     

Viable bacteria present within oral squamous cell carcinoma tissue

Despite increasing interest in the possible relationships between bacteria and the different stages of cancer development, the association of bacteria with cancer of the oral cavity has yet to be adequately examined. With that in mind, the primary objective of this study was to identify any bacterial species within oral squamous cell carcinoma tissue using a standard microbiological culture approach. At the time of surgery, a 1-cm3 portion of tissue was harvested from deep within the tumor mass using a fresh blade for each cut. Whenever possible, "superficial" portions from the mucosa overlying the tumor and nontumorous control specimens from at least 5 cm away from the primary tumor site were also obtained. Surface contamination was eliminated by immersion in Betadine and washing with phosphate-buffered saline. Each specimen was aseptically macerated and cultured on nonselective media under both aerobic and anaerobic conditions. Isolates were identified by 16S rRNA gene sequencing. Twenty deep-tissue specimens, 19 with corresponding superficial tissues and 12 with control tissues, were successfully processed. A diversity of bacterial taxa were isolated and identified, including several putatively novel species. Most isolates were found to be saccharolytic and acid-tolerant species. Notably, some species were isolated only from either the tumorous or nontumorous tissue type, indicating a degree of restriction. Successful surface decontamination of the specimens indicates that the bacteria detected were from within the tissue. A diversity of bacterial groups have been isolated from within oral squamous cell carcinoma tissue. The significance of these bacteria within the tumor warrants further study.     

Morphometric study of desmosomes from oral squamous cell carcinoma

The authors studied ultrastructural characteristics of desmosomes from oral squamous cell carcinoma, reporting the cellular differentiation and size of the desmosomes. The length of the desmosome profiles was measured with a Zeiss KS-300. The desmosomes were grouped according to their size and the tumor histological grading. Statistical analysis indicated a significant correlation (p < .001) between the size of the desmosomes and the histological grading group of the malignancy. The comparison of the desmosome size among the tumor histological grading groups also showed significant difference (p < .001). A multiple comparisons test indicated homogeneity in the size of desmosomes within the histological grading groups: 100% in the well differentiated, 95.2% in the moderately differentiated, and 50% in the poorly differentiated group. The preliminary data strongly suggest that the homogeneity of length of the desmosome profiles may be exploited for diagnostic strategies.