Roles of the matrix metalloproteinases in mammary gland development and cancer

Tissue remodeling is a key process involved in normal development, wound healing, bone remodeling, and embryonic implantation, as well as pathological conditions such as tumor invasion and metastasis, and angiogenesis. The degradation of the extracellular matrix that is associated with those processes is mediated by a number of families of extracellular proteinases. These families include the serine proteinases, such as the plasminogen-urokinase plasminogen activator system and leukocyte elastases, the cysteine proteinases, like cathepsin D and L, and the zinc-dependent matrix metalloproteinases (MMPs) [1]. Accumulating evidence has highlighted the central role of MMP-driven extracellular matrix remodeling in mammary gland development and breast cancer.     

Postoperative wound complications and systemic recurrence in breast cancer

Many factors involved in wound healing can stimulate tumour growth in the experimental setting. This study examined the relationship between wound complications and the development of systemic recurrence after treatment of primary breast cancer. One thousand and sixty-five patients diagnosed with operable primary invasive breast cancer between 1994 and 2001 were assessed for development of systemic recurrence according to whether or not a wound complication occurred after surgery, with a median follow-up of 54 months (range 15-119). There were 93 wound complications (9%). There was a statistically significant greater risk of developing systemic recurrence in patients with wound problems than those without (hazard ratio (HR) 2.87; 95% CI: 1.97, 4.18; P<0.0001). This remained in a multivariate analysis after adjustment for case mix variables, including Nottingham Prognostic Index (NPI) and oestrogen-progesterone receptor status (HR: 2.52; 95% CI: 1.69, 3.77; P<0.0001). In the good prognostic NPI group, 4 out of 27 patients (15%) with wound problems vs 11 out of 334 (3%) without wound problems developed systemic recurrence. The corresponding figures were 10 out of 35 (29%) vs 48 out of 412 (12 %) in the moderate prognostic group and 18 out of 29 (62%) vs 75 out of 199 (38%) in the poor prognostic group. In 29 patients NPI could not be calculated. Smokers at the time of diagnosis were more likely to develop metastatic disease than the non-smokers (HR: 1.50; 95% CI: 1.04, 2.15; P=0.03) after adjustment for other factors. The results suggest that patients with wound complications at primary surgery have increased rates of systemic recurrence of breast cancer.     

Study of matrix metalloproteinases and their inhibitors in breast cancer

An immunohistochemical study was performed using tissue microarrays and specific antibodies against matrix metalloproteinases (MMPs) 1, 2, 7, 9, 11, 13, 14, and their tisullar inhibitors (TIMPs) 1, 2, and 3. More than 2600 determinations on cancer specimens from 131 patients with primary ductal invasive tumours of the breast (65 with and 66 without distant metastasis) and controls were performed. Staining results were categorised using a score based on the intensity of the staining and a specific software program calculated the percentage of immunostained cells automatically. We observed a broad variation of the total immunostaining scores and the cell type expressing each protein. There were multiple and significant associations between the expression of the different MMPs and TIMPs evaluated and some parameters indicative of tumour aggressiveness, such as large tumour size, advanced tumour grade, high Nottinham prognostic index, negative oestrogen receptor status, peritumoural inflammation, desmoplastic reaction, and infiltrating tumoural edge. Likewise, the detection of elevated immunohistochemical scores for MMP-9, 11, TIMP-1, and TIMP-2, was significantly associated with a higher rate of distant metastases. The expression of MMP-9 or TIMP-2 by tumour cells, MMP-1, 7, 9, 11, 13, or TIMP-3 by fibroblastic cells, and MMP-7, 9, 11, 13, 14, TIMP-1, or TIMP-2 by mononuclear inflammatory cells, was also significantly associated with a higher rate of distant metastases.     

乳腺癌术后早期化疗与伤口愈合关系的临床观察

目的 总结乳腺癌术后早期化疗对伤口愈合的影响及适应证。方法 乳腺癌患者88例，均行简化根治术，术后选择性对其中53例患者行术后早期化疗对其临床资料进行总结分析。结果 术后早期化疗的53例乳腺癌患者的伤口临床观察30天,无1例发生皮下积液、伤口感染、皮肤坏死和伤口延迟愈合，伤口拆线在9～15天。 结论 乳腺癌术后早期化疗对伤口的愈合无明显影响。     

Polymorphisms of matrix metalloproteinases 1, 2, 3 and 9 and susceptibility to lung,breast and colorectal cancer in over 30,000 subjects

A variety of susceptibility genes have been associated with cancer but definitive conclusions have been difficult to draw partly hampered by the small number of subjects in each study. We undertook a comprehensive genetic meta‐analysis of all matrix metalloproteinase (MMP) genes investigated using an allelic‐association case–control model in the 3 major cancers of lung, breast and colorectal cancer. Electronic databases were searched until and including July 2008 for any MMP genetic association study in lung, breast and colorectal cancer. Odds ratio (OR) and 95% confidence intervals (CI) were determined for each gene disease association using fixed and random effect models. Twenty‐five studies addressing 5 polymorphisms in 4 genes were analyzed among 30,651 individuals (15,328 cases and 15,253 controls). The MMP‐1 nt‐1607 polymorphism was significantly associated with colorectal cancer in both the dominant (OR, 1.66; 95% CI, 1.14–2.42; p = 0.008) and recessive (OR, 1.59; 95% CI, 1.15–2.20; p = 0.005) models. MMP‐2^1306C→T (OR, 0.53; 95% CI, 0.40–0.72; p < 0.0001) and 735^C→T (OR, 0.65; 95% CI, 0.53–0.79; p < 0.0001) were significantly associated with protection against lung cancer. No association was found with the MMP 1, 2, 3 or 9 polymorphisms with breast cancer, MMP‐1, 3 or 9 with lung cancer or MMP‐2, 3 or 9 with colorectal cancer. There may be a genetic influence in the development of colorectal and lung cancer. Subjects with the MMP‐1 nt‐1607 polymorphism have an increased association with colorectal cancer. Those homozygous for either the MMP‐2/1306T or 735T allele may be at reduced risk of lung cancer, although the evidence base is small. © 2009 UICC     

High birth weight increases mammary tumorigenesis in rats

Epidemiological studies have investigated whether a high birth weight is associated with increased breast cancer risk, but the results remain inconclusive. This study was designed to determine whether high birth weight increases later susceptibility to carcinogen-induced mammary tumorigenesis in an animal model and to determine mechanisms mediating this association. Pregnant female Sprague Dawley rats were fed either a control or a high-fat diet during the extent of gestation. Maternal exposure to the high-fat diet increased pregnancy leptin levels and offspring's birth weight, but had no effect on pregnancy estradiol or insulin-like growth factor 1 levels. Changes in the offspring's mammary gland morphology and protein expression were assessed. The mammary epithelial tree of the high-birth-weight offspring was denser, contained more terminal end buds and exhibited higher number of proliferating cells. Further, their mammary glands expressed lower levels of ER-alpha, but higher levels of activated MAPK. No alterations in apoptosis were noted. High-birth-weight rats developed 7,12-dimethylbenz[a]anthracene-induced mammary tumors significantly earlier, and tumors grew larger than in the controls. The tumors in this group expressed higher levels of leptin receptor and activated Akt, and contained fewer apoptotic cells than those in the controls. Our results indicate that high birth weight is related to shortened latency to develop mammary tumors--perhaps reflecting an increase in activated MAPK levels and increased tumor growth--perhaps caused by a lower apoptotic response due to higher leptin receptor and activated Akt levels in the tumors.     

ras癌基因在人乳腺肿瘤中的表达

应用P_(21)蛋白单克隆抗体,对25例人乳腺癌的免疫组织化学检测显示,在乳腺浸润性导管癌组织中,有高水平的P_(21)表达,阳性率高达92％,与Querzoli等报道一致。29伊4乳腺良性病变的检测结果显示,细胞内P_(21)表达水平的高低与细胞增殖状态相关。临床对比分析提示乳腺癌中P_(21)的表达水平与瘤体大小有关但不能反映病人的淋巴结转移状态。上述结果表明,ras癌基因表达产物P_(21)蛋白,是一个与细胞增殖相关,可反映组织增殖状态的细胞生物学因子。     

High‐throughput proteomics of breast cancer interstitial fluid: identification of tumor subtype‐specific serologically relevant biomarkers

Despite significant advancements in breast cancer (BC) research, clinicians lack robust serological protein markers for accurate diagnostics and tumor stratification. Tumor interstitial fluid (TIF) accumulates aberrantly externalized proteins within the local tumor space, which can potentially gain access to the circulatory system. As such, TIF may represent a valuable starting point for identifying relevant tumor‐specific serological biomarkers. The aim of the study was to perform comprehensive proteomic profiling of TIF to identify proteins associated with BC tumor status and subtype. A liquid chromatography tandem mass spectrometry (LC‐MS/MS) analysis of 35 TIFs of three main subtypes: luminal (19), Her2 (4), and triple‐negative (TNBC) (12) resulted in the identification of > 8800 proteins. Unsupervised hierarchical clustering segregated the TIF proteome into two major clusters, luminal and TNBC/Her2 subgroups. High‐grade tumors enriched with tumor infiltrating lymphocytes (TILs) were also stratified from low‐grade tumors. A consensus analysis approach, including differential abundance analysis, selection operator regression, and random forest returned a minimal set of 24 proteins associated with BC subtypes, receptor status, and TIL scoring. Among them, a panel of 10 proteins, AGR3, BCAM, CELSR1, MIEN1, NAT1, PIP4K2B, SEC23B, THTPA, TMEM51, and ULBP2, was found to stratify the tumor subtype‐specific TIFs. In particular, upregulation of BCAM and CELSR1 differentiates luminal subtypes, while upregulation of MIEN1 differentiates Her2 subtypes. Immunohistochemistry analysis showed a direct correlation between protein abundance in TIFs and intratumor expression levels for all 10 proteins. Sensitivity and specificity were estimated for this protein panel by using an independent, comprehensive breast tumor proteome dataset. The results of this analysis strongly support our data, with eight of the proteins potentially representing biomarkers for stratification of BC subtypes. Five of the most representative proteomics databases currently available were also used to estimate the potential for these selected proteins to serve as putative serological markers.

Abbreviations

AUC

area under the curve

BC

breast cancer

DAA

differential abundance analysis

ER

estrogen receptor

FIF

fat interstitial fluid

Her2

human epidermal growth factor receptor 2

IHC

immunohistochemistry

LASSO

least absolute shrinkage and selection operator

MPs

microparticles

NIF

normal interstitial fluid

PgR

progesterone receptor

PPI

protein‐protein interaction

RF

random forest

TIF

tumor interstitial fluid

TILs

tumor infiltrating lymphocytes

TNBC

triple‐negative breast cancer

     

乳腺癌合并高血压患者术后伤口愈合不良分析

目的:分析乳腺癌合并高血压患者行改良根治术后伤口愈合不良的原因,探寻促进伤口愈合的方法。方法:随机选取我院2012年8月-2014年8月收治的76例乳腺癌患者的临床资料,按是否并发高血压分为两组,对照组无并发症37例;并发高血压组39例,对比两组患者伤口愈合情况。结果:对照组平均置管时间（19.1±7.0）天,并发高血压组平均置管时间（29.3±18.3）天,引流量增多。与对照组比较,并发高血压组患者伤口愈合迟缓,两组患者术后伤口愈合情况比较差异有统计学意义（P<0.01）。结论:并发高血压组乳腺癌患者行改良根治术后引流管留置时间较长,伤口愈合较迟缓。积极调控血压,维持血压平稳,术中术后正确医护,将有效促进患者伤口愈合,明显提高患者术后生活质量。     

Association of kallikrein expression in nipple aspirate fluid with breast cancer risk

Human kallikreins (hK) 2, 3, 6 and 10 are expressed in breast and prostate tissue. hK2 and hK3 (prostate-specific antigen, PSA) are used to screen for prostate cancer. hK6 and hK10 are downregulated in breast cancer compared to normal breast tissue. We demonstrated that levels of PSA in nipple aspirate fluid (NAF) are lower in women with breast cancer than in normal women. We hypothesize that the expression of hK2, 3, 6 and 10 are related and important in detecting breast cancer. The goals of this study are to determine the level of expression of kallikreins in NAF and serum, the association of hK2, 3, 6 and 10 in NAF, and the association of each of the kallikreins with breast cancer. In NAF from 275 women, hK3, 6 and 10 were detectable in >/= 90% and hK2 in 74% of samples analyzed. NAF levels were highest for hK6 and lowest for hK2, regardless of cancer and menopausal status. hK3 was detectable in 15/29 (52%) and hK2 in 0/29 serum samples collected from 6 women. hK2 and hK3 were concentrated in NAF vs. matched serum. The 4 kallikreins were associated with the exception of hK2 with hK6 or hK10. PSA levels were higher in normal pre- than postmenopausal subjects (but not women with breast cancer), whereas levels of hK2, 6 and 10 did not differ by menopausal status. hK2 and PSA were associated with both pre- and postmenopausal breast cancer; hK6 and 10 were not. hK2 and PSA were more associated with pre- than postmenopausal breast cancer. Using logistic regression, PSA and menopausal status provided the best model of breast cancer prediction, with a sensitivity of 91% and specificity of 39%. In conclusion, 4 kallikreins are expressed in NAF. hK2 and PSA, and hK6 and hK10 are highly associated. Higher premenopausal PSA levels suggest the influence of ovarian steroids. PSA shows the most promise in aiding in the early detection of breast cancer.     

Overexpression of matrix metalloproteinases and their inhibitors in mononuclear inflammatory cells in breast cancer correlates with metastasis-relapse

An immunohistochemical study was performed using tissue microarrays and specific antibodies against matrix metalloproteinase (MMP)-1, -2, -7, -9, -11, -13 and -14, tissular inhibitors of metalloproteinase (TIMP)-1, -2 and -3. More than 2600 determinations on cancer specimens from 131 patients with primary ductal invasive tumours of the breast were performed. To identify specific groups of tumours with distinct expression profiles the data were analysed by unsupervised hierarchical cluster analysis by each cellular type. We did not find well-defined cluster of cases for tumour cells or fibroblastic cells. However, for mononuclear inflammatory cells the dendogram shows a first-order division of the tumours into two distinct MMP/TIMP molecular profiles, designated group 1 (n=89) and group 2 (n=42). Matrix metalloproteinase-7, -9, -11, -13 and -14, and TIMP-1 and -2, were identified as showing significant high expression in group 2 compared with group 1. Multivariate analysis demonstrated that clustering for mononuclear inflammatory cells was the most potent independent factor associated with distant relapse-free survival (group 2: 5.6 (3.5-9.6), P<0.001). We identify a phenotype of mononuclear inflammatory cells infiltrating tumours, which is associated with the development of distant metastasis. Therefore, this finding suggests that these host inflammatory cells could be a possible target for inhibition of metastasis.     

Lower sensitivity of screening mammography after previous benign breast surgery

Few data are available on the effect of previous benign breast surgery on screening mammography accuracy. We determined whether sensitivity of screening mammography and tumor characteristics are different for women with and without previous benign breast surgery. We included a consecutive series of 317,398 screening mammograms of women screened between 1997 and 2008. During 2-year follow-up, clinical data, breast imaging, biopsy and surgery reports were collected from women with screen-detected or interval breast cancers. Screening sensitivity, tumor biology and tumor stages were compared between 168 women with breast cancer and prior ipsilateral benign breast surgery and 2,039 women with breast cancer but without previous ipsilateral, benign breast surgery. The sensitivity of screening mammography was significantly lower for women with prior surgery [64.3% (108/168) versus 73.4% (1,496/2,039), p = 0.01]. The concomitant increased interval cancer risk remained significant after logistic regression adjustment for age and breast density (OR = 1.5, 95% CI: 1.1-2.1). Comparing screen-detected cancers in women with and without prior breast surgery, no significant differences in estrogen receptor status (p = 0.56), mitotic activity (p = 0.17), proportions of large (T2+) tumors (p = 0.6) or lymph node positive tumors (p = 0.4) were found. Also for interval cancers, no differences were found in estrogen receptor status (p = 0.41), mitotic activity (p = 0.39), proportions of large tumors (p = 0.9) and lymph node positive tumors (p = 0.5) between women with and without prior breast surgery. We conclude that sensitivity of screening mammography is significantly lower in women with previous benign breast surgery than without, but tumor characteristics are comparable both for screen detected cancers and interval cancers.     

Mammographic appearances of mammary duct ectasia that mimic carcinoma in a screening programme

The aim of this study is to demonstrate the spectrum of mammographic appearances of mammary duct ectasia that mimic carcinoma in a breast cancer screening programme. Between February 1989 and March 1993, 40 003 women underwent screening mammography as part of the Western Australia Women's Cancer Prevention Unit screening programme. Fine needle aspiration or excisional biopsy was performed on 1437 women, and 12 cases of cytologically or histologically confirmed mammary duct ectasia were detected. A total of 14 mammographic abnormalities from 12 asymptomatic female patients were biopsied, and confirmed to represent mammary duct ectasia. The mammographic spectrum included eight areas of microcalcification (two of which were extensive), three spiculated masses and three lobulated, partially smooth masses. Five of these women showed no other mammographic stigmata of mammary duct ectasia in either breast. Additional features of mammary duct ectasia, including nipple retraction, retro-areolar duct dilatation or macrocalcification were identified in seven women. Mammographic features of mammary duct ectasia are frequently detected in asymptomatic women undergoing screening mammography and cause no diagnostic dilemma. Occasionally mammary duct ectasia will have a mammographic appearance that is indistinguishable from carcinoma, necessitating breast biospy. In this study 40% of those women with mammary duct ectasia that were submitted for biopsy had no other feature of mammary duct ectasia that could have suggested the pre-operative diagnosis.     

Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

Nidogen 1 (NID1) is a glycoprotein found in basement membranes involved in cross-linking collagen IV and laminin. The role of NID in breast cancer has only been evaluated in a small number of studies and the findings of these studies have been inconsistent. Our previous work revealed that highly tumorigenic murine mammary tumor cells express high levels of Nid1 while weakly tumorigenic mammary tumor cells express low levels of Nid1. To investigate Nid1, two stable knockdown lines were created, and Nid1 knockdown was confirmed at both the mRNA and protein level. Nid1 knockdown significantly reduced cell proliferation and migration/invasion and these reductions in proliferation and migration/invasion could be rescued by conditioned media containing NID1 protein. The reduced migration/invasion observed in the Nid1 knockdown cells was not associated with significant alterations in the epithelial gene Cdh1 or the mesenchymal genes Snai1, Snai2, Twist1, Twist2, Zeb1 and Zeb2. Therefore, suppression of Nid1 expression reduces proliferation and migration/invasion in claudin-low murine mammary tumor cells.     

Induction of matrix metalloproteinases MMP-1 and MMP-2 by co-culture of breast cancer cells and bone marrow fibroblasts

Two invasive breast cancer cell lines (MDA-MB-231 and BT-549) were found to be more adherent and have greater migratory capacity on bone marrow fibroblasts than three non-invasive cell lines (MCF-7, T47D and BT-483). Antibodies to the adhesion molecules CD44, E-cadherin, ICAM-1, and integrin chains 2, 3, 4, 5, 6, v, 1, 3 and 7 failed to inhibit breast cancer cell migration through bone marrow fibroblasts. Inhibitors of matrix metalloproteases, 1, 10-phenanthroline, Ro-9790, TIMP-1 and TIMP-2 were able to attenuate the migration of MDA-MB-231 cells through bone marrow fibroblast monolayers suggesting a role for these enzymes in the migration of breast cancer cells through bone marrow adherent layers. Co-culture of MDA-MB-231 cells and bone marrow fibroblasts resulted in augmentation of the levels of the matrix metalloproteases MMP-1 and MMP-2 in culture supernatants. Soluble factors produced by bone marrow fibroblasts were responsible for the increase in MMP-1 levels. However, maximal MMP-2 production was dependent on direct contract between the breast cancer cells and the bone marrow fibroblasts. Modulation of MMP production by cell–cell contact or soluble factors suggests a mechanism by which breast cancer cells can enhance their ability to invade the bone marrow microenvironment.     

不同方法预防乳腺癌术后皮下积液疗效的比较

目的：观察乳腺癌术中、术后不同处理方法对患者术后皮下引流量及皮下积液发生率的影响,探讨预防皮下积液发生的有效方法。方法：将72例乳腺癌患者随机分为3组,每组24例,分对照组、单纯加压包扎组、加压包扎联合术中应用医用生物蛋白胶组,比较3组患者术后皮下引流量、平均拔管时间以及皮下积液的发生率。结果：与对照组及单纯加压包扎组相比,加压包扎联合术中应用医用生物蛋白胶组的术后皮下引流量更少（P〈0.05）、术后带管时间更短（P〈0.05）、术后皮下积液的发生率更低（P〈0.05）。结论：术后胸部弹力绷带加压包扎联合术中应用医用生物蛋白胶是一种降低乳腺癌术后皮下积液发生的有效方法。     

Proteomic analysis of nipple aspirate fluid to detect biologic markers of breast cancer

The early detection of breast cancer is the best means to minimise disease-related mortality. Current screening techniques have limited sensitivity and specificity. Breast nipple aspirate fluid can be obtained noninvasively and contains proteins secreted from ductal and lobular epithelia. Nipple aspirate fluid proteins are breast specific and generally more concentrated than corresponding blood levels. Proteomic analysis of 1 microl of diluted nipple aspirate fluid over a 5-40 kDa range from 20 subjects with breast cancer and 13 with nondiseased breasts identified five differentially expressed proteins. The most sensitive and specific proteins were 6500 and 15 940 Da, found in 75-84% of samples from women with cancer but in only 0-9% of samples from normal women. These findings suggest that (1) differential expression of nipple aspirate fluid proteins exists between women with normal and diseased breasts, and (2) analysis of these proteins may predict the presence of breast cancer.     

A retrospective study of the effect of participation in screening mammography on the use of chemotherapy and breast conserving surgery

A population sample was obtained from the British Columbia (BC) Cancer Registry of all women diagnosed with a first breast cancer in 2002 who were resident in Greater Vancouver or Greater Victoria, BC. Information on treatment and prognostic factors were obtained from source records. The study group was linked to the records of the Screening Mammography Program of BC to identify screening histories on women prior to diagnosis. Logistic regression was used to determine the relationship between screening participation and treatment and to predict treatment use from prognostic factors. Fifteen hundred and eighty-nine women with breast cancer were included in the study and 1,071 (67%) had participated in screening prior to diagnosis: 786 (49%) had been screened within the 30 months prior to their diagnosis (regular participants). Breast conserving surgery (BCS) rates were higher (OR = 2.3, p < 0.001) and chemotherapy use lower (OR = 0.53, p < 0.001) among regular participants compared with nonparticipants after adjustment for age. A predictive model based on the distribution of prognostic factors between participants provided estimates of OR = 1.47 and OR = 0.54 for BCS and chemotherapy, respectively, and adjustment for self-selection changed the predicted values to OR = 1.16 and OR = 0.67, respectively. Participation in screening produced a considerable change in the use of chemotherapy but less on BCS use.     

Differences in predictors of cervical and breast cancer screening by screening need in uninsured Latino women

BACKGROUND: Latino women experience higher mortality for cervical cancer and lower 5-year survival for breast cancer than non-Latino White women. Adherence with screening recommendations can increase chances of survival, yet the factors that influence screening behaviors in uninsured women are not well documented. METHODS: Uninsured Latino women (N = 467) recruited in four US cities participated in the study. Logistic regression was used to model adherence to recommendations by screening type (cervical or breast cancer) and screening need (needs to obtain initial screening, overdue for rescreening, up-to-date with rescreening). RESULTS: Predictors differed by type of screening and screening need. Women who reported exposure to cancer education were more likely to have had a mammogram and to be up-to-date with Pap smear screening than women without such exposure. Women who were younger, had more than a sixth grade education, and/or had children were more likely to have had a Pap smear. Older women who had been in the US the longest were more likely to be overdue for a Pap smear. Women with incomes 5000 to 7000 were more likely to have obtained a mammogram. Regional differences were found with respect to mammography screening and maintenance behaviors. CONCLUSIONS: Exposure to cancer education is an important predictor of screenings among uninsured urban Latino women. The potential of creating educational interventions that can increase screening rates among women who evidence health disparities is encouraging. Recruitment strategies to reach women in need of screenings are provided.