Identification of TM9SF2 as a Candidate of the Cell Surface Marker Common to Breast Carcinoma Cells

OBJECTIVE We aimed identification of cell surface molecules, which might serve as diagnostic biomarkers or useful targets for therapies, in breast cancer. METHODS We developed unique DNA microarray coupled with spherical self-organizing map （sSOM） analysis to characterize cells and tissues by the cell surface markers. In the microarray 1,797 probes for human genes coding membrane bound proteins were spotted. With this microarray the gene expression profiles of eight breast carcinoma cell lines were compared to identify the genes that were commonly expressed in breast carcinomas but not in normal cells. RESULTS The gene expression profiles of sSOM from the eight breast carcinoma cell lines were successfully distinguished from that of normal breast tissue derived cells suggesting the presence of genes of interest, sSOMon the data extensively filtered revealed several candidate genes, of which expression was significant in carcinoma cells but low in normal cells. Finally, TM9SF2 was nominated through validations of PCR procedures together with CD24 and ErbB3, which are known breast carcinoma markers. TMgSF2 expression was further confirmed by immunological staining. Interestingly, TMgSF2 was found to be expressed in all the cell lines evaluated while CD24 and ErbB3 were not in all of the carcinoma cells, supporting their relationship in sSOM. Although physiological significance of TMgSF2 is unknown yet, siRNA treatment significantly inhibited the growth of MDA- MB-231 cells. CONCLUSION We propose TM9SF2 as a novel and useful diagnostic marker as well as a potential molecular target specific to breast carcinoma cells covering wide range of breast cancer.

Effect of A-L Tonic Capsule on DNA Content in Rat Experimental Hepatocarcinogenesis

The effects of A L tonic capsule on DNA content in rat experimental hepatocarcinogenesis induced by diethylnitrosamine (DENA) were observed. The experimental rats were divided into 4 groups. With exception of group D in which the rats were only administered with DENA, the rats in the groups A, B, C were previously, simultaneously and subsequently fed with A L tonic capsule respectively while they were administered with DENA. The DNA content of all rat livers was measured using automatic image analysis technique 20 weeks after administration of DENA. The results showed that the highest and lowest DNA contents were respectively seen in the groups D and A. There was significant difference in DNA contents between the groups A or B or C and D, and also between the groups A and B or C (both P <0.01). 4 components (4C) and ≥5C cells were predominant in the group D, while 2C cells were the minority. The number of 2C cells in the groups A, B, C was significantly higher than that in the group D, and the number of ≥5C cells in the groups A, B, C was markedly lower than that in the group D ( P <0.01). Also, there was very significant difference in the number of 2C and ≥5C cells between group A and B or C ( P <0.01). It was concluded that A L tonic capsule could effectively inhibit the increase of DNA content of hepatocytes and improve the distribution of DNA content in rat hepatocarcinogenesis, especially in group A.

Studies on the DNA content of breast carcinoma cells with neuroendocrine differentiation

OBJECTIVE: To make quantitative analysis of DNA content of breast cancer with neuroendocrine (NE) cells and its significance. METHODS: Using MIPS-III image analyzer, DNA content and 9 parameter measurements of the tumor nuclei were made in both NE positive (17) and negative (64) breast carcinomas. RESULTS: Out of 81 breast carcinomas, 17 cases were NE positive while 64 cases were NE negative. In the NE (+) cases, the integral optic density, mean optic density, DNA index, DNA stemlines peak, > 5c aneuploidy cells and the rate of aneuploidy cells were all lower than those in the NE negative breast carcinoma cases (P

COMPARATIVE PATHOLOGICAL AND FCM STUDIES ON THE PRIMARY AND METASTATIC CANCERS IN STOMACH AND BREAST

Pathological morphology,differentiation,cellular DNA content and cell Proliferation index(PI) were comparatively studied in the primary and their corresponding metastatic lesions of 54 cases of gastric and breast cancers- The results showed that differentiation and types of metastatic cancers were in accordance with their corresponding primary cancers in more than half cases(18/13) of stomach cancers,differences could be found in the other cases(16/34),and among them,10 were lower and 6 were higher differentially than their corresponding primary cancers.Similar results were found in breast cancer cases.Flow cytometry(FCM) analysis revealed that dramatic DNA Index(DI) differences between metastatic and their corresponding primary cancers existed in 6/34 of gastric and 4/18 of mammary cancers,among them 7 cases were higher than the primary while 3 were lower than the primary(DI).Similar results could also be found in PI analysis.All these suggested that metastasis of gastric and breast cancer was a very complicated process and metastatic cancer did not necessarily always show lower differentiation,higher DNA contents and DNA aneuploidy as well as higher cell proliferative rate.

DNA CYTOPHOTOMETRIC MEASUREMENTS OF CANINE STOMACH CARCINOGENESIS INDUCED BY N-ETHYL-N'-NITRO-N-NITROSO GUANIDINE

Measurement of DNA content and nuclear area by microspectrophotometry was performed in gastric carcinogenesis of three adult wolfdogs induced by N-ethyl-N’-nitro-N-nitrosoguanidine (ENNG). The mean values and standard deviations of nuclear DNA and area in different groups of lesions were as follows: normal gastric mucosal 0.03±2.30 AU and 28.76±5.85 μm^2; atrophic gastritis 12.04±3.34 AU and 28.69±8.02/μm^2; mild dysplasia 13.52±3.73 AU and 28.23±8.12μm^2; moderate dysplasia 20.88±4.57 AU and 47.58±10.74 μm^2; severe dysplasia 24.01±4.48 AU and 56.64±21.53μm^2; well-differentiated adenocarcinoma 33.07±9.38 AU and 72.99±15.57μm^2, respectively. The nuclear DNA content and nuclear area showed a significant difference statistically between normal gastric mucosa and gastric carcinoma (P【0.01). The nuclear area of gastric carcinoma increased with DNA content (r=0.73, P【0.01), and the distribution patterns of DNA content in the histogram showed that diploidy was decreased and polyploidy was increased in carcinoma. These findings indicate that DNA ploidy patterns and nuclear area could be a useful index in differentiated carcinoma and precancerous lesions.

大鼠乳癌发生过程中核DNA含量变化与cyclin D1、P53蛋白表达

目的 探讨乳腺癌发生过程中细胞核DNA含量变化、cyclinD1、P5 3蛋白表达情况及其相互关系。方法 雌性Wistar大鼠 12 0只 ,利用DMBA 2 0mg·(10 0g) - 1 灌胃 ,复制大鼠乳腺癌动物模型 ,观察正常乳腺、不典型增生乳腺和乳腺癌中细胞核DNA含量变化。应用SP免疫组化技术检测cyclinD1、P5 3蛋白在正常乳腺、不典型增生乳腺及乳腺癌中的表达。结果 在乳腺癌发生过程中 ,正常乳腺、不典型增生乳腺及乳腺癌中细胞核DNA含量增加 (P <0 .0 5 ) ;在不典型增生Ⅰ、Ⅱ、Ⅲ级乳腺组织中分别有 3 2 0、6 2 0、11 2 0有cyclinD1表达 ,7 9的乳腺癌Ⅰ级有cyclinD1的表达 ,1 4的乳腺癌Ⅱ级中cyclinD1有表达。P5 3蛋白在乳癌Ⅰ、Ⅱ、Ⅲ级中分别有 1 9、1 4、6 7有过度表达。结论 在大鼠乳腺癌发生过程中 ,细胞核DNA含量变化反映细胞增殖状态 ,cyclinD1过度表达参与肿瘤的发生过程 ,是肿瘤发生中的早期事件 ,而P5 3表达与肿瘤进展有关 ,是肿瘤发生的晚期事件 ,cyclinD1表达与P5 3蛋白表达呈负相关。

流式细胞计测定食管癌细胞 DNA 含量和临床及病理特性的关系

本文用流式细胞检测了36例食管癌切除标本的DNA含量,结果发现,DNA倍体水平差异较大,范围为1.68C～8.16C,大多数食管癌为非整倍体DNA含量,其中少数病人在同一标本中同时发现两种不同的非整倍体。而且,食管癌细胞的DNA含量和肿瘤病变长度、局部淋巴结转移、食管外侵犯以及病理分级有密切关系;以二倍体肿瘤预后较好,且病理分化较高。而非整倍体肿瘤则相反。

PCNA及DNA含量与胃癌预后关系的研究

作者利用免疫组化LSAB法和流式细胞分析技术（FCM）对41例进展期胃癌组织中增殖细胞核抗原（PCNA）的表达及细胞核DNA含量进行了测定。结果表明：PCNA标记指数及DNA非整信体检出率与胃癌病人术后5年生存率呈明显负相关（P＜0．05），与淋巴结转移呈正相关（P＜0．05）。DNA非整信体检出率与PCNA标记指数具有良好的相关性，二者均可用于判断胃癌预后，而二者同时运用则更有参考价值。

乳腺癌细胞DNA含量分析

本文应用流式细胞术(Flow Cytomotry简称FCM)测定41例乳腺癌细胞DNA含量,分析结果:DNA指数(DI)范围为0.9～2.91,其中二倍体肿瘤13例,占31.7％,异倍体肿瘤28例,占68.3％。DI和病理组织学分级呈正相关系,随着肿瘤组织学分级的增高,DI也升高。但是,在目前研究中,DI尚不反映临床分期。DI和预后的关系,共观察19例,发现二倍体肿瘤的预后好,生存期长,异倍体肿瘤的预后差,生存期短。本文应用针吸肿瘤细胞和切除标本新鲜组织块检测DNA含量,发现两种取材方法所得数值无显著差别。因此,认为针吸细胞应用FCM的DNA检查可作为术前辅助诊断方法,为治疗提供参考。

乳腺癌BRCA1蛋白表达与DNA定量分析的临床研究

目的 研究柳州地区人散发性乳腺癌BRCA1蛋白及mRNA表达与DNA含量的关系及意义。方法 应用S P免疫组化法及图像分析检测2 5例正常乳腺上皮和增生上皮及5 0例散发性乳腺癌组织中BRCA1蛋白表达及DNA情况。应用原位杂交技术检测5 0例乳腺癌中的2 0例乳腺癌组织BRCA1mRNA表达。结果 在乳腺正常上皮、增生上皮和癌中BRCA1表达及DNA含量差异均具有显著性(P <0 . 0 1) ;BRCA1表达与DNA含量呈负相关(P <0. 0 1) ;BRCA1在不同肿瘤大小和不同组织学类型中的表达差异无显著性(P >0 . 0 5 ) ,但在不同组织分级和有无淋巴结转移中的表达差异有显著性(P <0 . 0 5 )。DNA分析中,DI值和>5C %值总体分布在不同组织学类型中差异无显著性(P >0. 0 5 ) ,但在不同肿瘤大小、不同组织分级和有无淋巴结转移中差异有显著性(P <0 . 0 5 )。结论 乳腺上皮中DNA含量增加,BRCA1表达下降,对散发性乳腺癌的发生发展具有一定作用。检测BRCA1表达及DNA含量在散发性乳腺癌的早期诊断、判断生物学行为、预测预后中有重要意义。

乳腺癌Ki-67抗原和DNA倍体的相关性及临床意义

目的 探讨乳腺癌中 Ki- 6 7抗原表达和 DNA倍体的相关性及临床意义。 方法 采用 SP免疫组织化学方法和 DNA图像分析技术检测 130例乳腺癌组织中 Ki- 6 7抗原表达和 DNA倍体 ,结合临床病理资料进行分析。 结果 Ki- 6 7抗原表达率与乳腺癌 TNM分期、腋淋巴结转移以及 DNA非整倍体率之间均存在显著正相关关系 (P<0 .0 1)。 结论 Ki- 6 7表达和 DNA倍体可以反映乳腺癌的细胞增殖活性 ,联合检测两指标对评估乳腺癌的生物学行为有一定的参考价值。

DNA双链断裂修复蛋白在乳腺癌和乳腺纤维腺瘤中的表达

目的研究DNA双链断裂修复蛋白(Ku80、DNA-PKcs和ATM)在乳腺癌和乳腺纤维腺瘤组织中的表达,探讨3种蛋白在乳腺癌发生发展中的作用及相互关系。方法应用免疫组化SP法检测55例乳腺癌和14例乳腺纤维腺瘤组织中Ku80、DNA-PKcs和ATM蛋白的表达情况。结果Ku80、DNA-PKcs和ATM蛋白在乳腺癌患者中的阳性率分别为74.5%、54.5%和52.7%,在乳腺纤维腺瘤患者中的阳性率分别为92.9%、92.9%和71.4%;DNA-PKcs蛋白在乳腺癌组织中的表达显著低于乳腺纤维腺瘤组织(χ2=6.975,P=0.008),而Ku80和ATM的表达虽在乳腺癌组织中的表达亦低于乳腺纤维腺瘤组织,但差异无显著性意义(均P>0.05)。ATM和DNA-PKcs蛋白的表达在不同病理类型、肿块大小、临床分期和淋巴结转移状态的乳腺癌患者中差异无统计学意义(P>0.05);但Ku80的表达与患者的肿块大小和临床分期相关(P<0.01)。Spearman等级相关分析提示:在55例乳腺癌患者中,Ku80与DNA-PKcs的表达呈正相关(r=0.355,P=0.008);Ku80与ATM的表达亦呈正相关(r=0.325,P=0.015)。结论DNA-PKcs蛋白可能在乳腺癌的发生发展中起重要作用;Ku80可以促进乳腺癌细胞生长;Ku80与DNA-PKcs及ATM存在密切关系。

全自动DNA定量技术能提高支气管镜刷片中肿瘤细胞的阳性检出率

目的探讨DNA定量技术肺癌筛查和早期诊断中的应用价值。方法对181例肺部疾病患者进行纤维支气管镜取活检并刷检。活检组织经常规固定、石蜡包埋、HE染色后进行病理检查;留取部分刷取物制作刷片,常规固定、HE染色后进行常规细胞学检查,剩余刷取物放入麦克奥迪DNA倍体样本处理试剂中固定,悬滴法制作涂片,经Feulgen染色,采用Motic全自动细胞DNA定量分析系统进行细胞DNA定量分析。对比分析细胞DNA定量分析与传统支气管刷片的敏感度和特异度。结果采用全自动DNA定量技术检测纤维支气管镜刷检物中肿瘤细胞的灵敏度为87.30%、显著高于传统纤维支气管镜刷片的灵敏度52.38%,但特异度(80.5%)较其低。结论全自动DNA定量技术是一种敏感而特异的肺癌筛查和早期诊断技术,能提高纤维支气管镜刷检物中肿瘤细胞检出率。

乳腺浸润性导管癌DNA含量与组织学分级的关系

目的探讨DNA含量与乳腺浸润性导管癌组织学分级的关系。方法参照Bloom-Richardson和Elston的方法对72例浸润性导管癌按腺管形成、核的多形性和核分裂像计数进行组织学分级,应用图像分析法检测DNA倍体情况。结果在浸润性导管癌中,Ⅰ级(15例)、Ⅱ级(38例)、Ⅲ级(19例),DI值分别是1.08±0.29、1.37±0.20、2.30±0.53。与乳头状瘤相比,各组间差异具有显著性(P<0.01)。结论在浸润性导管癌中,DNA倍体和组织学分级、临床分期同样可作为较好的预后指标。

乳癌及癌前病变细胞核内DNA定量研究

应用显微分光光度计检测40例乳癌、癌旁单纯增生、异型增生及正常导管上皮细胞核内的DNA含量,结果表明:癌细胞核内DNA含量明显高于正常和癌旁上皮,癌旁异型增生的程度与DNA含量之间有着内在联系,多倍体的出现是细胞恶变的早期表现。局部细胞癌变时DNA含量明显增高,且高于高分化腺癌。作者认为检测细胞核内DNA含量对乳癌的诊断,癌及癌前病变的分级以及早期发现癌变趋势具有重要意义。

细胞DNA指数（倍体）图像分析估计直肠癌的预后

本文用自行设计组装的Ｍ１ＰＳ－Ｉ型自动图像分析仪对２１例直肠癌组织切片进行细胞核ＤＮＡ多数测定，并探讨其５项指标与患者预后的关系，结果发现：生存期≥５年和＜５年两组的（１）ＤＮＡ指数均值（２）分布宽度（ａ）ＤＮＡ主干系水平［（倍体，Ｃ）］间有显著性差异（ｐ＜０．０５）。（４）两组患者的ＤＮＡ类型有２／３是异借体，１／３是多倍体，仅１例为二俗体，本文６Ｃ峰位细胞是直肠癌５年生存期的分界值，＜６Ｃ者，预后好，≥６０者，顶后差。（５）按患者细胞群体ＤＮＡ分布中主干系峰位的ＤＩ（Ｃ）值，分为三种峰型，Ⅰ型峰预后好，Ⅱ型峰次之，Ⅲ型峰最差。图像细胞定量化学表明，此为直肠癌独立的预后变量，这对Ｄｕｋｅｓ系统是一个极有价值的补充。

层粘连蛋白、DNA含量及细胞核面积对肺癌患者预后的影响

采用免疫组化染色，流式细胞术及体视学测量技术对38例肺癌标本进行LN表达、DNA含量及核面积均值测定，用多因素生存分析法分析这三种指标对肺癌患者生存期的影响。结果提示：多因素生存分析表明DI及核面积均值可作为肺癌患者的预后判断指标。

增殖细胞核抗原表达及细胞核DNA含量在胃癌早期诊断中的意义

应用免疫组化ＬＳＡＢ法及流式细胞分析技术（ＦＣＭ），对１２例早期胃癌，１５例胃癌前病变（８例胃粘膜中～重度不典型增生，７例轻度不典型增生）及６例正常胃粘膜增殖细胞核抗原（ＰＣＮＡ）的表达及细胞核ＤＮＡ含量进行了测定。着重探讨了胃粘膜癌变过程中ＰＣＮＡ的表达及ＤＮＡ含量变化的意义，试图为胃癌的早期诊断提供客观依据。结果表明：ＰＣＮＡ标记指数（ＬＩ）随胃粘膜病变程度的加重呈递增趋势。中～重度不典型增生与早期胃癌之间无明显差异（Ｐ＞０．０５）；ＤＮＡ非整倍体检出率比较，早期胃癌明显高于胃粘膜轻度不典型增生组（Ｐ＜０．０１）。

食管鳞状细胞癌DNA倍体异质性及其与患者预后关系的研究

本研究采用多点取材的方法，应用流式细胞术对８０例中晚期食管鳞状细胞癌的瘤体内ＤＮＡ倍体异质性进行了研究。ＤＮＡ异倍体的检出率为８８．８％（７１／８０），ＤＮＡ指数范围为０．７７～１．７４。８０例标本中，３８例瘤体内表现为ＤＮＡ倍体异质现象（４７．５０％）。ＤＮＡ倍体异质性肿瘤患者的５年生存率（３４．２％）极显著地低于非异质性肿瘤患者（６４．３％）（Ｐ＜０．０２５）。文中对瘤体内ＤＮＡ倍体异质性的可能意义和发生机制进行了讨论。

尖锐湿疣伴异型增生及癌变的DNA含量动态观察与研究

目的 探讨尖锐湿疣从异型增生到癌变的DMA含量变化规律 ,了解伴有异型增生的尖锐湿疣的癌前意义。方法 应用JDICM A图象分析系统检测 34例伴有异型增生Ⅰ～Ⅲ级尖锐湿疣 ,9例伴鳞癌变尖锐湿疣和 2 5例单纯尖锐湿疣细胞DNA含量。结果 单纯尖锐湿疣、尖锐湿疣伴异型增生Ⅲ级 ,尖锐湿疣合并原位鳞癌及尖锐湿疣伴鳞癌变之间DNA含量依次递增 ,差异有显著性 (P <0 .0 5及P <0 .0 0 1 )。单纯尖锐湿疣基本为 2倍体 (2C) ,随着细胞异型程度的升级直至癌变 ,2C区的比值依次递减 ,3C～ 4C区的比值依次递增 ,差异有极显著性 (P <0 .0 1及P <0 .0 0 1 ) ,其中尖锐湿疣伴异型增生Ⅲ级细胞含有大于 5C的非整倍体细胞。结论 尖锐湿疣伴有异型增生Ⅲ级的细胞是一种异常增殖细胞 。