Stromal myofibroblasts predict disease recurrence for colorectal cancer.

PURPOSE: Myofibroblasts, which are specifically differentiated fibroblasts, are thought to play a central role in the desmoplastic reaction, a dynamic stromal change closely associated with cancer development. Although fundamental studies suggest that myofibroblasts may either facilitate or inhibit cancer progression, cumulative evidence supports their role in promoting tumor progression. The aim of this study was to assess the value of myofibroblasts in the cancer stroma as an indicator of disease recurrence after colorectal cancer surgery. EXPERIMENTAL DESIGN: Using computer-assisted image analysis, we quantified myofibroblasts in the cancer-associated stroma of 192 colorectal cancers using alpha-smooth muscle actin as a marker. RESULTS: The cancer-associated stroma contained various numbers of myofibroblasts (0.35-19.0%; mean, 5.55 +/- 3.85%). Tumors with abundant myofibroblasts were associated with shorter disease-free survival rate (P = 0.001) for stage II and III colorectal cancer. Multivariate analysis indicated that alpha-smooth muscle actin was a significant prognostic factor comparable with lymph node metastasis and superior to other tumor and stromal components, including histology of the tumor invasive front, peritumoral lymphocytic infiltration, and Crohn's-like lymphoid reaction. Moreover, colorectal cancers with synchronous liver metastasis generally displayed an active desmoplastic reaction, which was retained in the metastatic lesion to a similar extent. CONCLUSIONS: The results suggest that the abundance of myofibroblasts in cancer-associated stroma may be a useful indicator of disease recurrence after curative colorectal cancer surgery.     

Matrix metalloproteinase-2 expression in stromal tissues is a consistent prognostic factor in stage II colon cancer

For patients with stage II colon cancer, the usefulness of adjuvant chemotherapy remains controversial. Therefore, it is important to identify high-risk indicators. The biological prognostic factors for recurrence might allow further insight into the optimal treatment strategy for patients with node-negative disease. Matrix metalloproteinase-2 seems to be one of the essential factors for tumor invasion and lymph node metastasis. In this study, we analyzed the expression of cyclooxygenase-2 and matrix metalloproteinase-2 by immunohistochemical staining in 109 patients with stage II colon cancer. A positive correlation was observed between tumor cyclooxygenase-2 and tumor matrix metalloproteinase-2 expression ( P =  0.0006) and between tumor cyclooxygenase-2 and stromal matrix metalloproteinase-2 expression ( P <  0.0001). Stromal matrix metalloproteinase-2 expression was associated with disease-free survival ( P =  0.0095) and was shown to be an independent risk factor for recurrence by multivariate analysis. In addition, we carried out an invasion assay in vitro to investigate whether cyclooxygenase-2 and matrix metalloproteinase-2 affected the tumor-invasive potential of colon cancer cell lines. The invasion assay showed that every cancer cell line acquired invasive potential in coculture with stromal cell lines and the cyclooxygenase-2 inhibitor suppressed this phenomenon by downregulating the matrix metalloproteinase-2 expression of stromal cells. In conclusion, these findings suggest that matrix metalloproteinase-2 expression in stromal cells can be a high-risk indicator for recurrence in patients with stage II colon cancer. ( Cancer Sci 2009; 100: 852–858)     

Myofibroblasts correlate with lymphatic microvessel density and lymph node metastasis in early-stage invasive colorectal carcinoma

BACKGROUND: Recent studies have shown that the interactions between tumor cells and stromal cells are important in tumor development. A possible correlation between tumor-activated myofibroblasts, the main component cells of tumor stroma, and lymphatic microvessel density (LMVD) or other clinical parameters in carcinoma was investigated. MATERIALS AND METHODS: Immunohistochemical examination of alpha-smooth muscle actin and podoplanin were performed in 83 cases of early-stage invasive colorectal carcinoma. RESULTS: There was a good correlation between proliferation of myofibroblasts (PMpt) and LMVD (LMVDpt) in the peri-tumoral area (p = 0.0034). Increased PMpt was also associated with lymphatic invasion (p = 0.0051) and with lymph node metastasis (p = 0.011). However, proliferation of myofibroblasts in intra-tumoral (PMit) areas was not associated with these clinical parameters. CONCLUSION: Proliferation of myofibroblasts in peri-tumoral areas seem to play an important role in lymphangiogenesis, and is also associated with lymph node metastasis.     

Proliferation of myofibroblasts in the stroma of epithelial hyperplastic lesions and squamous carcinoma of the larynx

OBJECTIVES: The immunohistochemical phenotype, distribution and significance of proliferation of myofibroblasts in laryngeal epithelial hyperplastic lesions (EHL) and squamous carcinoma (SC) were analyzed. METHODS: Samples of 42 resected larynxes and 40 laryngeal biopsies of EHL and SC were included. Immunohistochemistry was performed using antibodies against vimentin, alpha-smooth muscle actin (SMA), desmin and leukocyte common antigen. RESULTS: Myofibroblasts were vimentin- and SMA-positive, and were found exclusively in SC, indicating that invasion beyond the basement membrane is necessary to evoke a myofibroblastic stromal reaction. We observed two patterns of stromal reaction in SC: one was characterized by a marked proliferation of myofibroblasts and desmoplasia, with scarce lymphocytic infiltration; this pattern tended to be associated with well- or moderately differentiated SC. The other was characterized by few myofibroblasts, weak desmoplasia, and dense lymphocytic infiltration; the latter pattern tended to be associated with moderately or poorly differentiated SC. The degree of myofibroblast proliferation was inversely related to the density of lymphocytic infiltration. Antibodies against SMA also stained stromal blood vessels, demonstrating a gradual increase of vessel density as the grade of EHL increased. CONCLUSIONS: Immunohistochemical analysis of myofibroblasts provides useful information on the phenotypic characteristics of the stroma in laryngeal EHL and SC, and can serve as an additional marker of invasion.     

A reduced COX-2 expression and a reduced number of pericryptal myofibroblasts are associated with depressed adenoma of the colon

The histogenesis of depressed adenoma of the colon has not been sufficiently investigated. Pericryptal myofibroblasts are stromal cells expressing smooth muscle actin, and are involved in the differentiation and multiplication of epithelial cells in the colonic epithelium. COX-2 has been reported to be involved in the development of colon adenoma. We studied the histogenesis of depressed adenoma of the colon by examining the relationship between the presence of pericryptal myofibroblasts and COX-2 expression. Twenty-one depressed adenomas of the colon that had been resected endoscopically between June 1998 and May 2003 (mild-moderate atypia; mean diameter, 6.7 mm) and 23 elevated adenomas that had been resected endoscopically in 2003 (mild-moderate atypia; mean diameter, 11.7 mm), were studied. We performed immunohistochemical staining using alpha-smooth muscle actin antibody to detect pericryptal myofibroblasts. We also performed immunohistochemical staining for Cox-2. Eighteen (78.3%) of the 23 elevated adenomas and six (28.6%) of the 21 depressed adenomas were positive for pericryptal myofibroblasts immunohistochemically, showing a significant difference (P<0.001). Seventeen elevated adenomas (73.9%) and eight depressed adenomas (38.1%) were positive for COX-2 expression (P=0.016). COX-2 expression was detected in the stroma, and the sites of COX-2 expression coincided with the sites of pericryptal myofibroblasts. The histogenesis of depressed adenomas differs from that of elevated adenomas. Our results suggest that a low number of pericryptal myofibroblasts and a low COX-2 expression are associated with depressed adenomas.     

314例Ⅰb~Ⅱa期宫颈癌预后及淋巴结转移危险因素研究

  目的  回顾性研究探讨早期宫颈癌的临床病理特点、预后影响相关因素及淋巴结转移的高危因素。  方法  选择1999年1月至2005年1月在中山大学肿瘤防治中心妇科住院治疗的、经病理确诊的314例早期(Ⅰb~Ⅱa期)宫颈癌临床病例资料进行回顾性分析, 分析影响其预后及淋巴结转移的高危因素。  结果  314例病例5年生存率为88.0%, 复发率为13.4%。单因素分析显示深肌层浸润、脉管内瘤栓、淋巴结转移为总生存时间不良因素(P < 0.05)。Cox回归分析显示淋巴结转移、深肌层浸润是预后的独立危险因素(P < 0.05)。盆腔淋巴结转移组数≥3组与盆腔淋巴结转移组数 < 3组的生存时间有统计学差异(P=0.032)。单因素分析示SCCAg、FIGO分期、肿瘤直径、深肌层浸润、脉管内瘤栓、宫旁组织浸润均与盆腔淋巴结转移有关(P < 0.05)。Logistic回归多因素分析: 治疗前SCCAg > 3 ng/mL(P < 0.001, OR=4.966)、深肌层浸润(P=0.001, OR=5.503)与盆腔淋巴结转移有关。  结论  淋巴结转移、深肌层浸润是宫颈癌预后的独立危险因素: 治疗前SCCAg > 3 ng/mL、深肌层浸润是盆腔淋巴结转移的独立危险因素。      

Distribution of laminin and fibronectin isoforms in oral mucosa and oral squamous cell carcinoma

The expression of laminin and fibronectin isoforms varies with cellular maturation and differentiation and these differences may well influence cellular processes such as adhesion and motility. The basement membrane (BM) of fetal oral squamous epithelium contains the laminin chains, alpha2, alpha3, alpha5, beta1, beta2, beta3, gamma1 and gamma2. The BM of adult normal oral squamous epithelium comprises the laminin chains, alpha3, alpha5, beta1, beta3, gamma1 and gamma2. A re-expression of the laminin alpha2 and beta2 chains could be shown in adult hyperproliferative, dysplastic and carcinomatous lesions. In dysplasia and oral squamous cell carcinoma (OSCC), multifocal breaks of the BM are present as indicated by laminin chain antibodies. These breaks correlate to malignancy grade in their extent. Moreover, in the invasion front the alpha3 and gamma2 chain of laminin-5 can immunohistochemically be found outside the BM within the cytoplasm of budding carcinoma cells and in the adjacent stroma. The correlation between the morphological pattern of invasive tumour clusters and a laminin-5 immunostaining in the adjacent stroma may suggest, first, that a laminin-5 deposition outside the BM is an immunohistochemical marker for invasion and second, that OSCC invasion is guided by the laminin-5 matrix. Expression of oncofetal fibronectins (IIICS de novo glycosylated fibronectin and ED-B fibronectin) could be demonstrated throughout the stromal compartment. However, the ED-B fibronectin synthesizing cells (RNA/RNA in situ hybridization) are confined to small stroma areas and to single stroma and inflammatory cells in the invasion front. A correlation of the number of ED-B fibronectin synthesizing cells to malignancy grade could not be seen. ED-B fibronectin mRNA-positive cells seem to be concentrated in areas of fibrous stroma recruitment with a linear alignment of stromal fibro-/myofibroblasts (desmoplasia). Double staining experiments (ED-B fibronectin in situ hybridization and alpha-smooth muscle actin immunohistochemistry) indicated that the stroma myofibroblasts are a preferential source of ED-B fibronectin. In conclusion, in OSCC, a fetal extracellular matrix conversion is demonstrable. Tumour cells (laminin alpha2 and beta2 chain) and recruited stromal myofibroblasts (oncofetal ED-B fibronectin) contribute to the fetal extracellular matrix milieu.     

Clinicopathological significance of stromal myofibroblasts in invasive ductal carcinoma of the breast.

The purpose of this study was to investigate the distribution of CD34-positive fibroblasts and alpha-smooth muscle actin (alpha-SMA)-reactive myofibroblasts in the stroma of benign and malignant breast lesions and, secondly, to determine whether the presence of stromal myofibroblasts is associated with some of the clinicopathological characteristics of patients with invasive ductal carcinoma. The presence of stromal CD34-positive fibroblasts and myofibroblasts was investigated (as defined immunohistochemically) in 8 normal breast tissue samples, 58 invasive ductal carcinomas, 9 ductal carcinomas in situ and 16 specimens with benign lesions of the breast (fibroadenomas, ductal hyperplasias). We further studied the correlations between the presence of stromal myofibroblasts with 7 clinicopathological parameters in 58 invasive ductal carcinomas. The results indicated that the stroma of normal breast tissues contained CD34-positive fibroblasts. All benign breast lesions exhibited stromal CD34-positive fibroblasts. In contrast, the stroma of ductal carcinomas showed a complete loss of CD34-positive fibroblasts. alpha-SMA expression in stromal fibroblasts (myofibroblasts) was not detected in normal tissue samples or benign lesions except in 1 case of fibroadenoma, whereas positive myofibroblasts were found in 44.4% of ductal carcinomas in situ and 56.9% of invasive breast carcinomas. Comparison of clinicopathological parameters between invasive ductal carcinomas with and without stromal myofibroblasts revealed significant differences in lymph node metastasis, high histological grade and high microvessel density. These results suggest that CD34 loss and the presence of myofibroblasts favor the diagnosis of breast carcinoma. In invasive ductal carcinoma, the presence of stromal myofibroblasts correlated significantly with pathological parameters associated with a poor prognosis.     

Stromal myofibroblasts and malignant transformation in a 4NQO rat tongue carcinogenesis model

This study assesses the correlation of changes in the density of stromal myofibroblasts to propagation of histopathologic alterations and proliferative activity of the epithelium in a rat 4NQO-induced tongue rat carcinogenesis model.Forty-three male Wistar rats were administered 0.001% 4NQO in drinking water for: 0 (n = 7, control), 7 (n = 4), 8 (n = 8), 14 (n = 6), 22 (n = 9), and 28 (n = 9) weeks, after which they were euthanized. Tongue sections were divided equally into anterior, middle and posterior thirds. Each third was given a histopathologic score (hematoxylin and eosin), ranging from normal, hyperplasia/hyperkeratosis, and escalating degrees of dysplasia to carcinoma, and analyzed by the point-counting method for density of epithelium-associated stromal myofibroblasts (SMA) and of proliferating epithelial cells (PCNA). Histopathologic changes significantly increased in severity (carcinoma) with duration of 4NQO administration (p < 0.001) in the posterior third of the tongue. The density of stromal myofibroblasts and proliferating epithelial cells was significantly higher in the posterior third of the tongue at 28 weeks compared to the other time points and locations (p < 0.001, p = 0.01, respectively). Significant correlations were found between occurrence of carcinoma and the increase in density of stromal myofibroblasts (p < 0.001) and of proliferating epithelial cells (p = 0.001) in the posterior third of the tongue. Increased density of stromal myofibroblasts was distinctively associated with the development of carcinoma but not with pre-malignant changes. Defining the mechanism of evolvement of carcinoma-associated stromal myofibroblasts is expected to further broaden our knowledge on the micro-environmental events occurring during the malignant transformation.     

Release of TGFβig-h3 by gastric myofibroblasts slows tumor growth and is decreased with cancer progression

Tumor progression has been linked to changes in the stromal environment. Myofibroblasts are stromal cells that are often increased in tumors but their contribution to cancer progression is not well understood. Here, we show that the secretomes of myofibroblasts derived from gastric cancers [cancer-associated myofibroblasts (CAMs)] differ in a functionally significant manner from those derived from adjacent tissue [adjacent tissue myofibroblasts (ATMs)]. CAMs showed increased rates of migration and proliferation compared with ATMs or normal tissue myofibroblasts (NTMs). Moreover, conditioned medium (CM) from CAMs significantly stimulated migration, invasion and proliferation of gastric cancer cells compared with CM from ATMs or NTMs. Proteomic analysis of myofibroblast secretomes revealed decreased abundance of the extracellular matrix (ECM) adaptor protein like transforming growth factor-β-induced gene-h3 (TGFβig-h3) in CAMs, which was correlated with lymph node involvement and shorter survival. TGFβig-h3 inhibited IGF-II-stimulated migration and proliferation of both cancer cells and myofibroblasts, and suppressed IGF-II activation of p42/44 MAPkinase; TGFβig-h3 knockdown increased IGF-II- and CM-stimulated migration. Furthermore, administration of TGFβig-h3 inhibited myofibroblast-stimulated growth of gastric cancer xenografts. We conclude that stromal cells exert inhibitory as well as stimulatory effects on tumor cells; TGFβig-h3 is a stromal inhibitory factor that is decreased with progression of gastric cancers.     

E-cadherin和CK19表达与甲状腺乳头状癌患者临床病理特征和复发的关系

目的:探讨上皮型钙黏附素（E-cadherin）和角蛋白19（CK19）与甲状腺乳头状癌（PTC）患者临床病理特征及复发的关系。方法:收集2012年7月至2015年6月期间在广元市中心医院行外科手术的PTC患者130例，随访患者复发情况。采用免疫组织化学染色法（IHC）检测所有PTC患者肿瘤组织和癌旁正常组织E-cadherin和CK19的表达。结果:肿瘤组织中E-cadherin阳性表达率为41.54%（54/130），明显低于癌旁组织，且与肿瘤腺体包膜浸润、淋巴结转移数量有关（P＜0.05）；而CK19阳性表达率为66.92%（87/130），明显高于癌旁正常组织，与肿瘤大小、腺体包膜浸润、TNM分期有关（P＜0.05）。E-cadherin在PTC组织中的表达与CK19呈负相关性（rk=-0.239，P=0.026）。14例（11.38%）患者术后复发。E-cadherin阳性表达者无病生存率高于阴性表达者（P＜0.05）。另外，CK19阳性表达者无病生存率低于阴性表达者（P＜0.05）。经多因素logistic回归分析，腺体包膜浸润、E-cadherin低表达和CK19高表达是术后复发的独立影响因素（P＜0.05）。结论:肿瘤组织中E-cadherin低表达和CK19高表达可作为PTC患者术后复发的预警标志物，有助于筛选术后复发的高危患者。     

Blockage of transdifferentiation from fibroblast to myofibroblast in experimental ovarian cancer models

Background  

Tumour stromal myofibroblasts can promote tumour invasion. As these cells are genetically more stable than cancer cells, there has been enormous interest in developing targeted molecular therapies against them. Chloride intracellular channel 4 (CLIC4) and reactive oxygen species (ROS) have been linked with promoting stromal cell transdifferentiation in various cancers, but little is known of their roles in ovarian cancer. In this study, we examined the functional roles that both CLIC4 and ROS play in the process of ovarian cancer cell-stimulated or TGF-β1 induced fibroblast-to-myofibroblast transdifferentiation. We also examine whether it is possible to reverse such a process, with the aim of developing novel therapies against ovarian cancer by targeting activated transdifferentiated myofibroblasts.     

The role of myofibroblasts at the tumor border of invasive colorectal adenocarcinomas

BACKGROUND: In order to elucidate the significance of myofibroblasts in invasive growth of colorectal adenocarcinomas, we examined the number of myofibroblasts at the tumor border of colorectal adenocarcinomas. METHOD: A total of 91 invasive colorectal adenocarcinomas were examined immunohistochemically using anti-alpha-smooth muscle actin (ASMA) and high-molecular-weight caldesmon (h-CD) antibodies; 25 carcinomas confined to the submucosa (sm carcinomas), 40 carcinomas confined to the muscularis propria (mp carcinomas) and 26 carcinomas invading the subserosa or adventitia (ss carcinomas). We considered ASMA-positive and h-CD-negative stromal cells as myofibroblasts. RESULTS: Twenty-seven (67%) of the 40 mp carcinomas and 25 (96%) of the 26 ss carcinomas had a small number of myofibroblasts at the tumor border facing the muscularis propria. CONCLUSIONS: Although direct evidence is lacking, there is a possibility that the further immediately vertical and radial invasion of carcinoma cells into the subserosa or adventitia is associated with a smaller number of myofibroblasts at the tumor border facing the muscularis propria in mp carcinomas, resulting in a low incidence of mp and a high incidence of ss carcinomas in the colorectum.      

Myofibroblasts at the tumor border of invasive gastric carcinomas: with special reference to histological type and tumor depth

In order to elucidate the role of alpha-smooth muscle actin (ASMA)-positive stromal cells, namely myofibroblasts (MFs), in invasive growth of gastric carcinomas (GCs), we examined the number of MFs at the deep border (DB) of GCs. In total 78 invasive GCs (48 intestinal type GCs and 30 diffuse type GCs) were examined immunohistochemically, and analyzed from the view point of tumor depth and histological type. In the intestinal type GCs examined, both the GCs confined to the submucosa and muscularis propria had none or a small number of MFs at the tumor border facing the submucosa (smDB) and muscularis propria (mpDB), respectively; whereas the GCs invading the subserosa had a moderate or large number of MFs at the tumor border facing the subserosa (ssDB) (p<0. 05). Regardless of tumor depth, the diffuse type GCs examined generally had none or a small number of MFs at the smDB, mpDB and ssDB, respectively. There is a possibility that intestinal type GCs invading the subserosa showing tubular and papillary structure may induce myofibroblastic transformation of gastric subserosal stromal fibroblastic cells.     

Collagenase-3 expression in breast myofibroblasts as a molecular marker of transition of ductal carcinoma in situ lesions to invasive ductal carcinomas

Collagenase-3 (matrix metalloproteinase 13; MMP-13), a protease originally identified in breast carcinoma, is characterized by a potent degrading activity against a wide spectrum of extracellular matrix proteins. The aims of this study were to localize and identify the MMP-13-expressing cells in invasive human breast carcinoma and to evaluate the role of MMP-13 in transition to invasive lesions by studying ductal carcinoma in situ (DCIS). We found expression of MMP-13 in stromal fibroblast-like cells in all 21 invasive ductal carcinomas studied and in 4 of 9 invasive lobular carcinomas. In most carcinomas, expression of MMP-13 was limited to small stromal foci in the tumor area. Combined in situ hybridization and immunohistochemistry showed coexpression of alpha-smooth muscle actin immunoreactivity and MMP-13 mRNA in myofibroblasts. In contrast, cytokeratin-positive cancer cells, alpha-smooth muscle actin-positive vascular smooth muscle cells, CD68-positive macrophages, and CD31-positive endothelial cells were all MMP-13 mRNA negative. In situ hybridization for MMP-13 in 17 DCIS lesions revealed expression in 10 cases. Immunohistochemical analysis of all DCIS cases identified microinvasion in 8 of the 17 lesions. Seven of the eight lesions with microinvasion were MMP-13 positive. Further analysis showed that MMP-13 expression was often associated with the microinvasive events. This particular expression pattern was unique for MMP-13 among other MMPs analyzed, including MMP-2, -11, and -14. We conclude that MMP-13 is primarily expressed by myofibroblasts in human breast carcinoma and that expression in DCIS lesions often is associated with microinvasive events. On the basis of these data, we propose that MMP-13 may play an essential role during transition of DCIS lesions to invasive ductal carcinomas.     

Tumour-derived TGF-beta1 modulates myofibroblast differentiation and promotes HGF/SF-dependent invasion of squamous carcinoma cells

The development of an altered stromal microenvironment is a common feature of many tumours including squamous cell carcinoma (SCC), and there is increasing evidence that these changes in the stroma, which include increased expression of proteases and cytokines, may actually promote tumour progression. A common finding is that stromal fibroblasts become 'activated' myofibroblasts, expressing smooth muscle actin and secreting cytokines, proteases and matrix proteins. We show that myofibroblasts are commonly found in the stroma of oral SCC and are often concentrated at the invasive margin of the tumour. Using oral SCC cells and primary oral fibroblasts, we demonstrate that tumour cells directly induce a myofibroblastic phenotype, and that this transdifferentiation is dependent on SCC-derived TGF-beta1. In turn, myofibroblasts secrete significantly higher levels of hepatocyte growth factor/scatter factor compared with fibroblast controls, and this cytokine promotes SCC invasion through Matrigel, a mixture of basement membrane proteins. This is the first time that this double paracrine mechanism has been demonstrated between squamous carcinoma cells and fibroblasts, and emphasises that cancer invasion can be promoted indirectly by the release of tumour-induced host factors from stroma.     

In vivo and in vitro invasiveness of a rat colon-cancer cell line maintaining E-cadherin expression: An enhancing role of tumor-associated myofibroblasts

In various cell systems, an inverse relationship was found between expression of E-cadherin, a molecule involved in the Ca²⁺-dependent homophylic cell-to-cell attachment of epithelial cells, and the capacity to invade extracellular matrix gels or normal tissues in vitro., DHD/K12/TRb (PROb) cells, maintained as a cell line derived from a rat colon carcinoma, homogeneously expressed. in vitro immunoreactive E-cadherin, which was functional as shown in cell dissociation-reassociation assays. PROb cells were found to be non-invasive in 3 different assays in vitro., However, tumors resulting from a s.c. injection of PROb cells into syngeneic BD-IX rats were invasive, although PROb cells maintained E-cadherin expression in the tumors. Cells from a freshly dissociated PROb tumor showed, not only PROb cells but also tumor-associated myofibrobfasts and were able to cross a Matrigel-coated filter. PROb tumors were indeed infiltrated by numerous myofibroblasts, mainly located at the invasive edge of the tumor. Cells from an established culture of tumor-infiltrating myofibroblasts were able to confer upon PROb cells invasiveness through Matrigel-coated filter or into chick-heart fragments. PROb cells maintained their capacity to express E-cadherin after myofibroblast-enhanced Matrigel invasion. Tumor-associated myofibroblasts, but not PROb cells, secreted a 72-kDa collagenase that could play a role in tumor-cell invasion. These results strongly suggest that cells from the tumor stroma, and more specifically myofibroblasts, may be involved in the invasiveness of epithelial tumor cells in vivo, even when E-cadherin expression prevents tumor-cell invasiveness in different in vitro assays.     

AJUBA在口腔鳞癌中的表达和意义及其相关机制

  目的  探讨AJUBA (ajuba LIM protein)基因在口腔鳞癌(oral squamous cell carcinoma,OSCC)中的表达及其对OSCC细胞增殖、迁移能力的影响。  方法  采用qPCR和免疫组织化学方法检测OSCC和癌旁组织中AJUBA的表达情况,分析AJUBA与OSCC临床病理参数之间的关系。通过MTT实验、划痕实验及Transwell实验探究AJUBA对OSCC细胞增殖、迁移和侵袭的影响。West-ern blot法检测AJUBA对OSCC细胞Snail/E-cadherin信号通路相关蛋白表达的影响。  结果  AJUBA在OSCC组织中显著高表达,与T分期、肿瘤分化、淋巴结转移和复发相关(P < 0.05),且其表达提示不良预后。敲降AJUBA后,OSCC细胞的增殖、迁移和侵袭能力受到抑制。Western blot结果显示AJUBA能够促进Snail的表达,抑制E-cadherin的表达。  结论  AJUBA在OSCC组织中高表达,可能通过Snail/E-cadherin信号通路参与OSCC的增殖和侵袭。      

Individual tumorigenesis pathways of sporadic colorectal adenocarcinomas are associated with the biological behavior of tumors

Clinicopathologic features of sporadic colorectal adenocarcinomas were compared using integrated data from 244 patients subjected to curative resection. Individual steps in the tumorigenesis pathway, that is, adenomatosis polyposis coli (APC), Wnt-activated, base excision repair mutations, mismatch repair defects, RAF-mediated, transforming growth factor (TGF)-b-suppressed, bone morphogenic protein (BMP)-suppressed, and p53 alterations, were examined in terms of genetic and epigenetic changes, as well as protein expression. Genetic and molecular alterations of right colon cancers were distinct from those of left colon and rectal cancers. Rectal cancers showed the attenuated phenotype of left colon cancers. Tumors most frequently displayed either TGF-b- or BMP-suppressed alterations (81.2%), followed by RAF-mediated alterations (78.6%), and mismatch repair defects (38.4%), constituting a total of 24 integrated pathways. Tumors lacking APC mutations or carrying the RAF alteration (V600E) were frequently associated with lymphovascular invasion and lymph node metastasis ( P <  0.05). Poorly differentiated or mucinous adenocarcinomas were generally associated with high level microsatellite instability, Axin2 suppression, TGF-b1 or BMPR1A suppression, loss of heterozygosity of D18S46 or D18S474 , and absence of base excision repair mutations ( P <  0.0001–0.05). Early tumor recurrence was significantly correlated with lack of APC mutations ( P =  0.036). Moreover, tumors that concurrently displayed APC/Wnt-activated, TGF-b/BMP-suppressed, and p53 alterations were significantly predisposed to early recurrence ( P =  0.026). Our data clearly indicate that particular steps or pathways of colorectal tumorigenesis are closely associated with characteristic clinicopathologic features that, in turn, determine biological behavior, such as tumor growth, invasion, and recurrence. ( Cancer Sci 2008; 99: 1348–1354)