Cell Localization and Redistribution of the 67 kD Laminin Receptor and α6βl Integrin Subunits in Response to Laminin Stimulation: An Immunogold Electron Microscopy Study

The 67 kDa laminin receptor (67LR), one of several cell surface laminin-binding proteins, is involved in the interactions between cancer cells and laminin during tumor invasion and metastasis. A 37 kDa polypeptide (37LRP), previously identified as the 67LR precursor, is abundantly present in the cytoplasm and has been implicated in polysome formation. To better understand the cellular localization of the 67LR and its precursor, transmission electron microscopic studies of human melanoma A2058 cells were carried out using immunogold labeling and a variety of antibodies: (a) affinity purified antibodies directed against 37LRP cDNA-derived synthetic peptides; (b) anti-67LR monoclonal antibodies raised against intact human small cell lung carcinoma cells; and (c) monoclonal antibodies against the subunits of the integrin laminin receptor, α6β1. Double-labeling immunocyto-chemistry revealed that anti-67LR monoclonal antibodies as well as anti-37 LRP antibodies recognized antigens that were localized in the cytoplasm in electron dense structures. As expected, cell membrane labeling was also observed. Surprisingly, α6 and β1 integrin subunits were detected in the same cytoplasmic structures positive for the 67LR and the 37LRP. After addition of soluble laminin to A2058 cells in suspension, the number of labeled cytoplasmic structures increased especially in the vicinity of the plasma membrane, and were exported onto the cell surface. Neither fibronectin nor BSA induced such an effect. The data demonstrate that the 67 LR and the 37 LRP antibodies detect colocalized antigens that are in cytoplasmic structures with α6β1 integrin. These laminin binding protein rich structures could potentially form a supply of receptors that are exported to the surface upon exposure of the cells to laminin, with a consequent increase in the number of binding sites for the ligand. This system could define a mechanism through which cancer cells modulate their interaction with laminin.     

Targeting multiple tyrosine kinase receptors with Dovitinib blocks invasion and the interaction between tumor cells and cancer-associated fibroblasts in breast cancer

A constitutive and dynamic interaction between tumor cells and their surrounding stroma is a prerequisite for tumor invasion and metastasis. Fibroblasts and myofibroblasts (collectively called cancer associated fibroblasts, CAFs) often represent the major cellular components of tumor stroma. Tumor cells secret different growth factors which induce CAFs proliferation and differentiation, and, consequently, CAFs secrete different chemokines, cytokines or growth factors which induce tumor cell invasion and metastasis. In this study we showed here that CAFs from breast cancer surgical specimens significantly induced the invasion of breast cancer cells in vitro. Most interestingly, the novel multiple tyrosine kinase inhibitor Dovitinib significantly blocked the CAFs-induced invasion of breast cancer cells by, at least in part, inhibition of the expression and secretion of CCL2, CCL5 and VEGF in CAFs. Inhibition of PI3K/Akt/mTOR signaling could be responsible for the effects of Dovitinib, since Dovitinib antagonized the promoted phosphorylated Akt after treatment with PDGF, FGF or breast cancer cell-conditioned media. Treatment with Dovitinib in combination with PI3K/Akt/mTOR signaling inhibitors Ly294002 or RAD001 resulted in additive inhibition of cell invasion. This is the first in vitro study to show that the multiple tyrosine kinase inhibitor has therapeutic activities against breast cancer metastasis by targeting both tumor cells and CAFs.     

The RUNX1 transcription factor is expressed in serous epithelial ovarian carcinoma and contributes to cell proliferation,migration and invasion

Previously, we have identified the RUNX1 gene as hypomethylated and overexpressed in post-chemotherapy (CT) primary cultures derived from epithelial ovarian cancer (EOC) patients, when compared with primary cultures derived from matched primary (prior to CT) tumors. Here we show that RUNX1 displays a trend of hypomethylation, although not significant, in omental metastases compared with primary EOC tumors. Surprisingly, RUNX1 displayed significantly higher expression not only in metastatic tissue, but also in high-grade primary tumors and even in low malignant potential tumors. The RUNX1 expression levels were almost identical in primary tumors and omental metastases, suggesting that RUNX1 hypomethylation might have a limited impact on its overexpression in advanced (metastatic) stage of the disease.

Knockdown of the RUNX1 expression in EOC cells led to sharp decrease of cell proliferation and induced G₁ cell cycle arrest. Moreover, RUNX1 suppression significantly inhibited EOC cell migration and invasion. Gene expression profiling and consecutive network and pathway analyses confirmed these findings, as numerous genes and pathways known previously to be implicated in ovarian tumorigenesis, including EOC tumor invasion and metastasis, were found to be downregulated upon RUNX1 suppression, while a number of pro-apoptotic genes and some EOC tumor suppressor genes were induced.

Taken together, our data are indicative for a strong oncogenic potential of the RUNX1 gene in EOC progression and suggest that RUNX1 might be a novel EOC therapeutic target. Further studies are needed to more completely elucidate the functional implications of RUNX1 and other members of the RUNX gene family in ovarian tumorigenesis.     

Role of the extracellular matrix in epithelial morphogenesis

The extracellular matrix (ECM) plays an essential role in organizing tissues, defining their shapes or in presenting growth factors. Their components have been well described in most species, but our understanding of the mechanisms that control ECM remodeling remains limited. Likewise, how the ECM contributes to cellular mechanical responses has been examined in few cases. Here, I review how studies performed in C. elegans have brought several significant advances on those topics. Focusing only on epithelial cells, I discuss basement membrane invasion by the anchor cell during vulva morphogenesis, a process that has greatly expanded our knowledge of ECM remodeling in vivo. I then discuss the ECM role in a novel mechanotransduction process, whereby muscle contractions stimulate the remodeling of hemidesmosome-like junctions in the epidermis, which highlights that these junctions are mechanosensitive. Finally, I discuss progress in defining the composition and potential roles of the apical ECM covering epidermal cells in embryos.     

AN ASSESSMENT OF THE POTENTIAL IMPACT OF ALIEN INVASIVE VEGETATION ON THE GEOMORPHOLOGY OF RIVER CHANNELS IN SOUTH AFRICA

Summary

Invasion of the riparian zone by alien vegetation is recognised as a serious problem in many areas of South Africa. Vegetation is a dynamic component of river channels. It is an important control variable affecting channel form whereas the flow and sediment regime influences vegetation growth. Wherever alien vegetation invades the riparian zone it can be expected that there will be some impact on the physical structure of the riparian habitat. This paper reviews the effect of riparian vegetation on channel processes and channel form and discusses the implications of the invasion of riparian zones by alien vegetation. Woody species in particular are seen as having a significant potential for inducing channel modification, whilst their removal could lead to significant channel instability and mobilisation of sediment. The need for further research into the impact of alien vegetation on the geomorphology of South African river channels is stressed.     

Species-specific impact of introduced largemouth bass Micropterus salmoides in the Groot Marico Freshwater Ecosystem Priority Area,South Africa

Largemouth bass Micropterus salmoides are among the world's 100 worst invaders and negatively affect aquatic biodiversity in many regions worldwide. In South Africa there is a paucity of empirical studies describing their impacts. The impact of M. salmoides on the fish community in the Groot Marico River catchment, an otherwise near-pristine river ecosystem and a freshwater ecosystem priority area, was assessed from surveys conducted in 2012. Fish presence and abundance were enumerated using multiple survey techniques, and their association with key habitat variables and the presence or absence of M. salmoides were assessed. A total of 14 native fish species were recorded, besides introduced M. salmoides, which occupied the majority of the mainstem and several tributaries downstream of barriers to upstream movement. Canonical correspondence analysis showed that only one native species, the Marico barb Barbus motebensis, had a negative spatial association with M. salmoides. Assessment of relative distributions showed this species to be excluded from M. salmoides-invaded river reaches, whereas the other native species were not visibly affected by the invader. This species-specificity of the impact of M. salmoides indicates that their impacts in South African streams may be dependent on predator-naiveté of prey.     

Effects of exotic guava (Psidium guajava L.) invasion on soil properties in Limpopo,South Africa

Many studies in South Africa have examined the impacts of alien plants on ecosystems, but none have assessed the impact of guava (Psidium guajava L.) invasion on soil properties. In this study, soils underneath guava-invaded sites were assessed to determine if they had different soil physico-chemical properties (pH, P, C, N, Na, K, Ca, Mg, moisture, penetration resistance, infiltration and water repellency) as compared to soils underneath uninvaded sites. Comparisons were made from three different sites over three autumn months. Results show that soil pH was significantly (p < 0.005) higher underneath uninvaded than guava-invaded sites. Soil P was three times higher underneath guava-invaded as compared to invaded sites. The soils collected underneath guava-invaded sites had a significantly (p < 0.001) higher moisture content and were less compact but more repellent than soils from the uninvaded sites. Infiltration rate was significantly (p < 0.001) higher in the uninvaded than the guava-invaded sites. The study concludes that guava invasion alters some soil properties, thus creating favourable conditions for its growth and making it potentially more invasive. From a management standpoint, guava removal is encouraged; however, given guava's socio-economic importance more research on cost and benefits is required.     

The roles of carbonic anhydrases IX and XII in cancer cell adhesion,migration, invasion and metastasis

The main function of carbonic anhydrases (CAs) in cancer cells is the pH regulation through a conversion of H₂O and CO₂ to H⁺ and HCO₃⁻. However, the data of in vitro and in vivo studies have demonstrated that transmembrane isoforms of CA IX and CA XII are involved in various steps of cancer cell migration, invasion and metastasis. According to literature, inhibition of these CAs can affect the expression of multiple proteins. Some scientific groups have reported the possible interactions between CA IX and E-cadherin–catenin system, CA IX and integrins, CA IX, CA XII and ion transporters, which all are highly involved in cell-to-cell adhesion, the formation of membrane protrusions and focal adhesions. Nevertheless, CA IX and CA XII have a high impact on tumour growth and metastases formation. The data discussed in this review are quite recent. It highly support the role of CA IX and CA XII in various cancer metastasis processes through their interactions to other invasion proteins. Nevertheless, all findings show the great potential of these CAs in the context of research and application in clinical use.     

Scavenging behaviour and size-dependent carcass consumption of the black bullhead (Ameiurus melas)

This study examined the size-dependent scavenging behaviour of black bullheads Ameiurus melas under laboratory conditions, using common bleak Alburnus alburnus and pumpkinseed Lepomis gibbosus carcasses. Video camera observations showed that the activity of A. melas was higher at night, but substantial daytime activity was also recorded. Larger A. melas were more active than their smaller conspecifics, especially at night. All size classes exhibited a well-defined sequence of consuming different parts of the carcasses independent of size, but larger individuals tended to consume carcasses more efficiently. Carcasses of the softer-bodied A. alburnus were consumed more readily than those of the bonier L. gibbosus, independent of size. This scavenging behaviour of A. melas might facilitate the invasion success of the species.     

Age,growth and reproductive traits of invasive goby Taenioides cirratus in the Chaohu Lake,China

The eel goby Taenioides cirratus (Blyth, 1,860) is a small fish inhabits muddy bottoms of brackish-water in the Indo-West Pacific. It has invaded many inland freshwater lakes in China, such as the Chaohu Lake, Gaoyou Lake and Nansi Lake, and its population increased rapidly in these freshwater lakes in recent years. The age, growth and reproductive traits of T. cirratus invading the Chaohu Lake were studied. A total of 482 specimens (210 females, 204 males and 68 juveniles) with total length (TL) ranging from 9.4 to 20.6 cm were collected using the benthic fyke nets at monthly intervals from March 2018 to February 2019. The sagittal otolith was used for age determination. Monthly variation of marginal increment ratio indicated that the annual forming of opaque band on sagittal otolith was completed during March and April. For both sexes, only four (from 0+ to 3+ years) age groups were observed and 1+ and 2+ years age individuals dominated the population. Back calculated length at age showed males grew faster than females. Both sexes reached maturity at 1+ year age and the TL at first maturity (TL₅₀) was 12.6 cm for females and 11.9 cm for males. Monthly variation of gonado-somatic index indicated that the spawning occurred from May to August. The fecundity ranged from 967 ova to 5,114 ova, with a mean of 3,205 ova. Our study provides a comprehensive data on the key life history traits of T. cirratus for the first time.