MDA-MB 231

Top PDF MDA-MB 231:

Studies on Focal Adhesion Kinase in human breast cancer cell MDA MB 231

Studies on Focal Adhesion Kinase in human breast cancer cell MDA MB 231

AIM: 1) To study the participation of Focal Adhesion Kinase (FAK) in regulation of Breast Cancer cell mi- gration in relation with MMP-9 and other signaling proteins; 2) To study the effect of some natural prod- ucts on FAK. METHODS: Cell culture, Western Blot, Immunoprecipitation, Immunocytochemistry, Zymogra- phy, SiRNA transfection, RT-PCR, Real-Time PCR. RESULTS: For our study on FAK, we selected inva- sive Breast Cancer cell line MDA-MB-231 and trea- ted the cells with Fibronectin (FN). Treatment of FN was found to increase FAK expression, phosphoryla- tion (Tyr 397). FAK was found to be involved in re- gulation of breast cancer cell migration and MMP-9 expression, activity. Fibronectin increases association of FAK with integrin α5β1, Paxillin, Actin, ERK, PI3K and localization at Focal Adhesion sites. FAK was found to be involved in modulation of ERK and PI3K phosphorylation. Moreover, FAK signal was found to be transduced through ERK and PI3K, which modulate MMP-9 and thereby cell migration. CON- CLUSION: FAK expression, phosphorylation and pro- cessing are induced in response to Cell-ECM interac- tions. Integrin α5β1 is involved in FN induced FAK phosphorylation. FAK is a potent regulator of MMP-9 expression and activity. FAK is involved in regulation of ERK and PI3K phosphorylation. ERK and PI3K are involved in FAK regulated MMP-9 expression & activity. FAK regulates MMP-9 expression and activ- ity and thereby migration of human breast cancer cell. By the regulation of FAK, cell attachment and migra- tion may be regulated by Curcumin, ATRA or EGCG treatment. It may be concluded that invasive poten- tial of breast cancer cells may be modulated by regu- lation of FAK.
Show more

14 Read more

Citral induced apoptosis in MDA MB 231 spheroid cells

Citral induced apoptosis in MDA MB 231 spheroid cells

signaling and downregulation of Wnt signaling related tumor suppressor genes such as APC, indicate higher levels of self-renewal and dyregulated proliferation of these cancer cells [22, 26]. A previous review has shown that drug targeting on aberrant Wnt signaling pathway can favour cancer treatment outcome [27]. For example, Niclosamide that blocked Wnt co-receptor LRP6 sup- pressed the growth of Wnt-driven MDA-MB-231 and T- 47D breast cancer cells [28]. In this study, we found that mRNA expression of Wnt destruction complex Axin, APC and CK1 were upregulated, while Wnt co-receptor LRP6 was downregulated in MDA-MB-231 spheroids by citral treatment in a dosage dependent manner. These activities are may lead to the phosphorylation of β - catenin and target the phospho β -catenin to proteosomal degradation, which essentially suppressed the Wnt/ β -ca- tenin signaling pathway and expression of its target gene, cyclin D1. These results have proposed that citral treatment may regulate the growth of MDA-MB-231 spheroids through downregulation of Wnt signaling and its downstream cell cycle target gene, cyclin D1. How- ever, further studies must be performed to validate the interaction of citral treatment with the components of the Wnt signaling pathway.
Show more

10 Read more

The effect of laser irradiation on the viability of human breast cancer cell, MDA-MB-231

The effect of laser irradiation on the viability of human breast cancer cell, MDA-MB-231

This study compared the effects of different sources of laser phototherapy on the cell viability of the in vitro human breast cancer cell lines. Laser phototherapy is used in the breast cancer clinical treatment, despite the limited safety information of laser irradiation effect on the cancer cell behavior. This study contributed on the development of guidelines for safer laser usage in treating breast cancer and minimising the possibility of activating postmastectomy lymphedema. Cancer cell viability and morphology were studied in this research. Human breast cancer (MDA-MB-231) cell lines were cultured for 24 hours in CO 2 incubator and irradiated with different laser sources and number of pulse. The
Show more

6 Read more

Mechanisms underlying the growth inhibitory effects of the cyclo oxygenase 2 inhibitor celecoxib in human breast cancer cells

Mechanisms underlying the growth inhibitory effects of the cyclo oxygenase 2 inhibitor celecoxib in human breast cancer cells

a likely candidate for the angiogenic response observed in several tumors, including mammary tumors [36,56-58]. To explore the role played by COX-2 inhibitors in angiogenesis, we used both in vitro and in vivo model systems. Aggressive breast epithelial cells are known to differentiate into tubules when cultured on growth factor reduced Matrigel. This phe- nomenon is known as vasculogenic mimicry. Its presence has been reported in inflammatory breast cancer patients and is associated with reduced 5-year survival and higher percent- age of recurrence [59]. Shirakawa and coworkers [40] sug- gested a connection between vascular mimicry and angiogenesis, based on the existence of blood flow in the vas- cular channels. When plated on growth factor reduced Matrigel, human breast cancer cell lines have the unique ability to form tubular channels. We showed that the more aggres- sive MDA-MB-231 cells generate channels more efficiently and in higher numbers than do the less aggressive MDA-MB- Figure 6
Show more

14 Read more

Cyanidin-3-o-glucoside directly binds to ER 36 and inhibits EGFR-positive triple-negative breast cancer

Cyanidin-3-o-glucoside directly binds to ER 36 and inhibits EGFR-positive triple-negative breast cancer

Cy-3-glu is the most widespread glycoside class of anthocyanin pigments in vegetables and fruits (Supplementary Table S1) [19]; the contents of Cy-3-glu is usually more than 10-100 times higher than delphinidin- 3-glucoside, petunidin-3-glucoside, peonidin-3-glucoside, pelargonidin-3-glucoside and malvidin-3-glucoside. To verify the efficacy of Cy-3-glu against BC, several cell lines representing different clinical subtypes of BC were examined. We treated MDA-MB-231 cells with low dose Cy-3-glu (5 and 10 μM) for a longer time (7 d) and found that the growth of MDA-MB-231 cells was significantly inhibited (p = 0.022) at 7 d (Supplementary Figure S1A). In order to study the mechanism and shorten the experiment time, we use the higher doses of Cy-3-glu (150 and 500 μM) and set up the time points at 24 and 48 h, respectively. We observed no significantly additional cytotoxicity at 48 h with higher dose (150 and 500 μM) of Cy-3-glu treatment (Supplementary Figure S1B). Treatment with both doses, the 150 and 500 μM of Cy-3- glu, significantly inhibited TNBC cell growth, as shown in Figure 1. Specifically, MDA-MB-231 (p < 0.001 at 48 h), MDA-MB-436 (p < 0.001 at 24 h and 48 h; 500 μM) and BT20 cells (p < 0.5 at 24 h and p < 0.001 at 48 h; 500 μM). Actually, a greater growth inhibitory effect was observed on MDA-MB-231 cells (Figure 1A and 1B), and this may due to different sensitivities of individual cell lines.
Show more

19 Read more

Gigantol inhibits Wnt/β catenin signaling and exhibits anticancer activity in breast cancer cells

Gigantol inhibits Wnt/β catenin signaling and exhibits anticancer activity in breast cancer cells

Fig. 5 Gigantol reduces the mRNA levels of Wnt target genes in breast cancer cells. a-d MDA-MB-231 (a and b) and MDA-MB-468 (c and d) cells were treated with the specified amounts of gigantol for 24 h. RNA was extracted and then reverse-transcribed into cDNA. Prepared cDNA was then subjected to real-time PCR analysis to detect the mRNA expression of Axin2 (a and c) and Survivin (b and d). Gigantol treatment significantly reduced the mRNA level of Axin2 and Survivin compared with DMSO treated control cells, * p < 0.05

8 Read more

Clinicopathological and functional significance of RECQL1 helicase in sporadic breast cancers

Clinicopathological and functional significance of RECQL1 helicase in sporadic breast cancers

RECQL1 depletion in breast cancer cells: On-Target plus SMARTpool small interfering RNAs (siRNAs) against RECQL1 (NM_032941), and non-targeting control (CTL) were purchased from Dharmacon (catalog nos. L-013597-00-0005 and D-001810-10-05, respectively). We have previously established the specificity of the siRNA pool (5). All siRNA transfections (in MCF-7, MDA-MB-231 and MDA-MB-468 breast cancer cells) were performed by reverse transfection at a final concentration of 20 nM using Lipofectamine RNAiMAX (Invitrogen, catalog no. 13-778-075) as instructed by the manufacturer. Stable shRNA-mediated knockdown of RECQL1 in MDA-MB-231 cells was achieved using a lentiviral system (26). Briefly, lentivirus particles were produced by cotransfecting 293T cells with the pLKO.1 lentiviral shRNA expression vector containing the RECQL1 targeting sequence (5”-GAGCTTATGTTACCAGTTA-3”) or the gene encoding Luciferase (5”- ACGCTGAGTACTTCGAAATGT-3”) with the packaging plasmids psPAX2 and pM2D.G; and used to transduce MDA-MB-231 cells, followed by selection with puromycin (8 µg/ml). All cells were cultured in a humidified atmosphere containing 5% CO 2 at 37 o C and routinely
Show more

62 Read more

Original Article DLC-3 suppresses cellular proliferation, migration, and invasion in triple-negative breast cancer by the Wnt/β-catenin pathway

Original Article DLC-3 suppresses cellular proliferation, migration, and invasion in triple-negative breast cancer by the Wnt/β-catenin pathway

Abstract: Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype. Our study investigated the functional role of DLC-3 in TNBC. The expression of DLC-3 was assessed by immunohistochemistry in TNBC to evaluate the clinicopathologic significance of DLC-3. Recombinant lentiviral vectors encoding the DLC-3 gene were constructed for transfection into MDA-MB-231. Real-time qPCR and western blot analysis were employed to evaluate the expression of DLC-3, β-catenin, GSK-3β and c-myc in DLC-3-transfected cells. Moreover, cell proliferation assays, cell colony formation assays, and cell migration and invasion assays were performed to elucidate the role of DLC-3 in TNBC development and progression. Our data revealed that DLC-3 was downregulated in TNBC, and its expres- sion level was associated with lymph node status and differentiation grade in breast cancer. Both real-time qPCR and western blot analyses showed that the DLC-3 gene and protein were overexpressed in the DLC-3-transfected MDA-MB-231 cells. In addition, the expression of GSK-3β was upregulated and the expression of β-catenin and c- myc gene was downregulated in the DLC-3-transfected cells. Furthermore, DLC-3 overexpression inhibited cell pro- liferation, colony formation, migration, and invasion in vitro. DLC-3, functioning as a tumor-suppressor gene, inhibits cell growth and invasion in TNBC, possibly through regulation of the Wnt/β-catenin signaling pathway.
Show more

9 Read more

Original Article Botulinum neurotoxin type A inhibits synaptic vesicle 2 expression in breast cancer cell lines

Original Article Botulinum neurotoxin type A inhibits synaptic vesicle 2 expression in breast cancer cell lines

MCF-10A, MDA-MB-231, MDA-MB-453 and T47D cell lines were donated by the Instituto Nacional de Cancerología, Secretaría de Salud in Mexico City. The MDA-MB-231 and MDA- MB-453 cell lines were cultured with DMEM (Sigma) and the T47D cell line with RPMI (Gibco). All culture media were supplemented with 5% bovine fetal serum and 1% antibiotic/antimy- cotic solution, and cells were incubated at 37°C with 5% CO 2 atmosphere. Every four days the cells were washed with sterile PBS and fresh medium was added, a procedure that was fol- lowed until 90% confluence was reached. Botulinum neurotoxin type A (BoNTA) and de- velopmental treatment
Show more

8 Read more

MicroRNA-454 may function as an oncogene via targeting AKT in triple negative breast cancer

MicroRNA-454 may function as an oncogene via targeting AKT in triple negative breast cancer

To determine the function of miR-454 on radiation- induced apoptosis in TNBC cells, the caspase-Glo 3/7 assay kit was used by measuring caspase 3/7 activity, and apoptotic related protein was examined by west- ern blot. As shown in Fig. 3, miR-454 inhibited the cas- pase 3/7 activity in both MDA-MB-231 (Fig.  3a) and MDA-MB-468 (Fig. 3b) cells at dose dependent manner. The expression of BAX and Bcl-2 was altered in MDA- MB-231 (Fig.  4c, d) and MDA-MB-468 (Fig.  4e, f) after upregulation of miR-454.

10 Read more

The effect of laser irradiation on survivability of breast canser cells sensitised with gold nanoparticles

The effect of laser irradiation on survivability of breast canser cells sensitised with gold nanoparticles

In this study, breast cancer cells MDA-MB-231, MDA-MB-468, MDA-Kb2 and normal cell from Chinese human ovary, CHO. All cells are cultured in Faculty of Bioscience and Medical Engineering, UTM and used as a sample to interact with non-ionized radiation. Gold nanoparticle (AuNPs) was fabricated by pulse laser ablation in liquid (PLAL) process and will be sterilized by several treatment techniques including ethanol, autoclaving, and ultraviolet exposure. The fabricated AuNPs will be characterized using Energy-filtered transmission electron microscopy (EFTEM) and the interested concentration and size will be verified. Kr2 Excimer, and Nd:YAG laser with tuneable wavelength including fundamental, and second harmonic generation were utilized as non-ionization sources. Energy density, and exposed duration for each of the source radiation will verified. The cell survivability rates after exposure were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) cell viability assay to observe the effect of cancer cell before and after radiation. Cell changes in morphology will be observed by inverted microscope.
Show more

32 Read more

Glutaminolysis drives membrane trafficking to promote invasiveness of breast cancer cells

Glutaminolysis drives membrane trafficking to promote invasiveness of breast cancer cells

and measured the appearance of degraded foci (which correspond to active invadopodia) underneath the cells. Consistent with previous reports, addition of the MMP inhibitor GM6001 com- pletely opposed degradation of gelatin by MDA-MB-231 cells (Fig. 5c). Importantly, withdrawal of glutamine or addition of LY95 also strongly opposed gelatin degradation, and addition of glutamate completely restored the ability of glutamine-starved cells to digest the fluorescent gelatin matrix (Fig. 5c). Finally, we used second harmonic generation (SHG) microscopy to deter- mine the amount of fibrillar collagen in organotypic plugs con- taining MMTV-PyMT cells and found that they degrade the ECM to generate areas with reduced fibrillar collagen content (Fig. 5d). CRISPR-mediated disruption or pharmacological blockade of GRM3 using LY95 significantly opposed the ability of MMTV- PyMT cells to reduce the amount of fibrillar collagen in orga- notypic plugs indicating that GRM3 is required for breast cancer cells to efficiently degrade the ECM in a 3D microenvironment (Fig. 5d).
Show more

15 Read more

<p>Estradiol promotes the progression of ER+ breast cancer through methylation-mediated RSK4 inactivation</p>

<p>Estradiol promotes the progression of ER+ breast cancer through methylation-mediated RSK4 inactivation</p>

Human breast epithelial HBL-100 cells incorporating SV40 gene and breast cancer ER+ cell line MCF-7, ER-negative (ER-) cell lines MDA-MB-231 and MDA-MB-453 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). MCF-7 cells were cultured in Eagle ’ s Minimum Essential Medium (Gibco, MA, USA); MDA-MB -231 and MDA-MB-453 cells were cultured in Leibovitz ’ s L-15 Medium (Gibco); HBL-100 cells were maintained in RPMI-1640 medium (Gibco). To make the complete growth medium, 10% fetal bovine serum (FBS; Gibco) was added to all kinds of cell culture mediums.
Show more

10 Read more

A novel HMGA1-CCNE2-YAP axis regulates breast cancer aggressiveness

A novel HMGA1-CCNE2-YAP axis regulates breast cancer aggressiveness

To further investigate whether CCNE2 is a direct target of HMGA1 in vivo, we performed chromatin immunoprecipitation (ChIP) assays in MDA-MB-231 cells. In addition of testing the promoter region we assayed also two more upstream regions, because from an inspection to the 5’ flanking region of the CCNE2 gene we observed that at -1.8 kb and at -8.6 kb there are two long regions of 3.0 kb and of 2.0 kb, that we named AT-rich region 1 (AT-RR1) and AT-rich region 2 (AT- RR2), with an unusual AT-rich composition, 62% and 55% respectively. A MatInspector analysis indicated the presence of several HMGA1 binding sites in these two regions and, in addition, the MAR-Wiz tool indicated in the more distant one (AT-RR2) a Scaffold/Matrix- Attachment-Region (S/MAR) suggesting the implication of these two putative regulatory elements in the regulation of the CCNE2 gene (Supplementary Figure S1e). ChIP experiments indicate that HMGA1 binds to the promoter region and, intriguingly, even more to AT-RR1 and AT- RR2 (Figure 1e), suggesting that HMGA1 could act through these two regulatory regions as well. Altogether therefore these results demonstrate that HMGA1 regulates CCNE2 expression by binding to the CCNE2 gene.
Show more

15 Read more

Long non-coding RNA SNHG22 facilitates the malignant phenotypes in triple-negative breast cancer via sponging miR-324-3p and upregulating SUDS3

Long non-coding RNA SNHG22 facilitates the malignant phenotypes in triple-negative breast cancer via sponging miR-324-3p and upregulating SUDS3

For the sake of searching the functions of SNHG22 in TNBC, we firstly wondered whether its expression changed in malignant conditions. We discovered that SNHG22 expression was extremely strong in TNBC cell lines relative to MCF-10A cells, particularly in MDA-MB-231 and MDA-MB-468 cells (Fig.  1a). There- fore, we implemented loss-of-function experiments in MDA-MB-231 and MDA-MB-468 cells who had higher original SNHG22 expression to investigate the influ- ence of SNHG22 on TNBC cell function. As observed in Fig.  1b, SNHG22 expression in these two cells was vis- ibly weakened after transfection of two shRNAs aiming at SNHG22. Expectedly, it was displayed that the colony formation ability and EdU-positive cell proportion were both declined after silencing SNHG22 (Fig. 1c, d), indi- cating that knockdown of SNHG22 evidently impaired TNBC cell proliferation. By contrast, we discovered that the relative caspase-3 activity was intensified in two TNBC cells when SNHG22 was inhibited (Fig.  1e), indi- cating that SNHG22 deficiency could expedite cell apop- tosis. Then the apoptosis-accelerating impact of SNHG22 suppression in TNBC cells was further evidenced by flow cytometry analysis (Fig.  1f). Moreover, we unveiled the restrained migration and invasion of the two TNBC cells in response to SNHG22 deficiency (Fig.  1g, h). Overall, SNHG22 was highly expressed in TNBC cells and facili- tated TNBC cell proliferation and motility.
Show more

12 Read more

Metabolic profiling of triple-negative breast cancer cells reveals metabolic vulnerabilities

Metabolic profiling of triple-negative breast cancer cells reveals metabolic vulnerabilities

We profiled pool sizes of 43 central carbon metabolites of subconfluent TNBC cell lines in exponential growth phase. In addition, we quantified pool size changes follow- ing treatment with small molecule inhibitors of the RTKs MET and EGFR. Both MET and EGFR were prominently expressed in the assayed cell lines. Hierarchical clustering of metabolic profiles of TNBC cell lines reveals molecular heterogeneity between the TNBC mesenchymal-like and basal-like subtypes (Fig. 1). Pool size measurements showed common clusters of low TCA cycle and elevated amino acid metabolites of mesenchymal-like MDA-MB- 231 and Hs578 which were distinct from the basal-like MDA-MB-468 and HCC70 cell lines (Fig. 1a, Additional file 4: Table S3). Drug perturbations of amino acid pool sizes demonstrated similar response of mesenchymal-like subtype MDA-MB-231 and Hs578 cell lines to both, INC280 or erlotinib, treatment (Fig. 1b). Clusters of each subtype and cell line were well separated by metabolic profiles and drug responses showing that each subtypes had major similarities but each breast cancer cell line also had distinct components. The TCA cycle organic acid α-ketoglutaric acid is significantly reduced upon INC280 treatment with p values below 0.05 for all tested TNBC cell lines. Similarly, TCA cycle and central carbon metabo- lites aspartic acid, fumaric acid, and malic acid are signifi- cantly reduced upon erlotinib treatment with p values
Show more

14 Read more

Suberoylanilide Hydroxamic Acid as a Potential Therapeutic Agent for Human Breast Cancer Treatment

Suberoylanilide Hydroxamic Acid as a Potential Therapeutic Agent for Human Breast Cancer Treatment

Since different doses of the compound may induce different cell cycle arrest, we further de- termined the cell cycle alteration profiles as a function of increasing doses of SAHA. Cells were treated with SAHA at concentrations ranging from 0.5 to 10 M for 24 and 48 hr and analyzed for cell cycle distributions (Fig. 4). In MCF7 cells, SAHA induced G1 arrest at the growth inhibitory low concentrations ( 2 M), and both G1 and G2/M arrest at high concen- trations ( 2 M), after 24 or 48 hr treatment. The percentage of cells in G2/M phase in- creased in a dose-dependent manner. As more cells accumulated at G2/M phase, less cells were at G1 phase. However, the G1-S bound- ary was still blocked, as seen by the dramatic reduction in S-phase cells. In MDA-MB-231 cells, at 24 hr, SAHA induced G1 and G2/M arrest at concentrations ranging from 1 to 10 M. At 48 hr, SAHA induced G1 arrest. No sig- nificant G2/M arrest was induced at this time point. In MDA-MB-435 cells, as in MCF7 cells, SAHA induced G1 arrest at low concentrations ( 2 M), and both G1 and G2/M arrest at high concentrations ( 2 M) at 24 and 48 hr. The percentage of cells in G2/M phase also in- creased in a dose-dependent manner.
Show more

18 Read more

Therapy response testing of breast cancer in a 3D high-throughput perfused microfluidic platform

Therapy response testing of breast cancer in a 3D high-throughput perfused microfluidic platform

The difference between 2D and 3D response is quite striking as cells in 3D generally show slower proliferation rates compared to 2D culture, and are expected to be less sensitive to anti-mitotic agents. However, Huyck et al. also showed higher sensitivity of MDA-MB-231 cells embed- ded in collagen to the thymidine synthesis inhibitor 5- fluoruracil [43]. Drug response in 3D might be further tuned by varying the composition of the ECM [17]. While initial growth factor concentrations in various ECM gels may impact survival of seeded cells, after a day or so of culture, growth factor contribution to cancer cell growth rate and viability following drug treatment is likely deter- mined by the far higher concentration of growth factors originating from serum in cell culture media.
Show more

11 Read more

Wogonoside inhibits invasion and migration through suppressing TRAF2/4 expression in breast cancer

Wogonoside inhibits invasion and migration through suppressing TRAF2/4 expression in breast cancer

Tumor metastasis accounts for 90% of cancer-associated deaths, in which invasion plays a critical role in metastasis [27]. During invasion, tumor cells firstly lose cell-cell junc- tions, subsequently degrade, remodel, and adhere to the surrounding ECM and eventually migrate through ECM to the distance sites [28]. Therefore, wound healing assay, transwell invasion assay and cell adhesion assay were used to measure the anti-migration, anti-invasion and anti- adhesion effect of wogonoside in vitro. We found that wogonoside could inhibit invasion and migration in TNF- α-induced MDA-MB-231, MDA-MB-435 and BT-474 cells. Meanwhile, the abnormal expression of MMP-9, MMP-2, vimentin and CD44v6 in cancer cells would lead to de- creased adhesion, enhanced migration and invasion. Thus, Wogonoside inhibited metastasis-related protein expression to block TNF-α-induced metastatic process. Twist1 is a master regulator of morphogenesis, which can induce EMT to facilitate breast tumor metastasis [6]. We found that wogonoside reduced the mRNA and protein expression of Twist1 in TNF-α-induced MDA-MB-231 and MDA-MB- 435 cells, which indicated that the anti-metastatic effect of wogonoside in breast cancer was dependent of Twist1 expression.
Show more

14 Read more

Development and immunochemical evaluation of a novel chicken IgY antibody specific for KLK6

Development and immunochemical evaluation of a novel chicken IgY antibody specific for KLK6

In particular, sensitivity of IgYs for detection of endogenous KLK6 and lack of crossreactivity with other KLKs and unre- lated proteins was assessed by analyzing whole cell lysates and supernatants isolated from MDA-MB-468 breast can- cer cell line that expresses the KLK5, KLK6 and KLK10 proteins. The MDA-MB-231 KLK6-non-expressing cell line was used as a negative control for KLK6 but a positive con- trol for KLK1 [32]. In addition, cDNA-transfected MDA- MB-231 cells with stably reconstituted KLK6 expression were included in the analysis [18]. The IgYs could detect the endogenous (secreted and intracellular) KLK6 protein produced by MDA-MB-468 and MDA-MB-231 cells stably transfected with the KLK6 cDNA (Figure 3C) and no other proteins were detected indicating high specificity of IgYs for detection of endogenous KLK6 by Westerns. On the other hand, complete absence of crossreactivity of the anti- KLK6 IgYs with endogenous KLK5 and KLK10 or any other KLK-unrelated proteins could be demonstrated in lysates and supernatants of MDA-MB-468. Also, lack of crossreactivity with KLK1 was shown in MDA-MB-231 cells (Figure 3C). If KLK10 was detected in MDA-MB-468
Show more

8 Read more

Show all 3081 documents...