Migration Of Human Prostate Cancer Cells Research Articles

Biochemical evaluation of selected grape varieties (Vitis vinifera L.) grown in Jordan and in vitro evaluation of grape seed extract on human prostate cancer cells

A chemical and biological approach was used for quality assessment of 16 different eating grape varieties collected in Jordan. A validated LC-ESI-MS/MS method was used for quantitative analysis of resveratrol in grape skin and grape seed extracts. Grape skin extracts showed higher resveratrol content than that of seeds. The antioxidant activity and total phenolic content of seed, skin, and whole berry extracts of the grape varieties were evaluated using the DPPH radical scavenging assay and Folin–Ciocalteu colorimetric method, respectively. Seed extracts showed the highest total phenolic content and antioxidant activity. The in vitro effects of Golden Scatt grape seed extract on human prostate cancer cell migration and colony-genic potential were assayed. Golden Scatt seed extract modulated the colony formation potential and inhibited the migratory potential of prostate cancer cells in a dose-dependent manner.

Downregulation of TRPM7 suppressed migration and invasion by regulating epithelial-mesenchymal transition in prostate cancer cells.

Metastasis is a leading cause of death in patients with prostate cancer (PCa). Transient receptor potential channel 7 (TRPM7) functions as a Mg2+/Ca2+-permeable channel as well as a protein kinase that regulate various cellular processes including cell adhesion, migration and survival. However, the function of TRPM7 in metastasis of PCa remains largely unknown. Microarray analysis suggested that calcium signaling pathway was significantly altered in metastatic PCa tissues, compared with benign prostatic hyperplasia tissues. Bioinformatics analysis using microarray data and database for annotation, visualization and integrated discovery database revealed altered genes involved in calcium signaling pathway were significantly upregulated in TRPM7 deficiency PCa cells, which was also confirmed by experimental verification. Therefore, we aim to investigate the role of TRPM7 in human PCa cell migration and invasion as well as the underlying mechanisms. We observed that TRPM7 was upregulated in PCa cells and tissues compared with prostate hyperplasia cells and tissues. Further investigations suggested that TRPM7 deficiency suppressed migration and invasion of distinct PCa cell lines while overexpression of TRPM7 increased migration of PCa cells. In addition, knockdown of TRPM7 in PCa cells reversed the epithelial-mesenchymal transition (EMT) status, accompanied by downregulation of MMPs and upregulation of E-cadherin. Taken together, our study indicated that downregulation of TRPM7 could inhibit migration and invasion via reversing EMT status in PCa cells.

Effects of upregulatioon of trophoblast cell-surface antigens 2 on migration, invasion and epithelial-mesenchymaltransition of human prostate cancer cells

Objective To explore the effects of over-expression of trophoblast cell-surface antigens 2(TROP-2)gene on the migration and invasion of human prostate cancer cells and the action mechanism. Methods The TROP-2 gene was cloned into the eukaryotic expression vector pcDNA3. 1, and TROP-2 over-expression of eukaryotic gene expression plasmid was constructed. All PC-3 cells were divided into three groups: the control group(Con-A), empty vector control group(Con-B)and over-expression group TROP-2(pcDNA3. 1-TROP-2).After PC-3 cells in three groups were transfected with phosphate buffer(PBS), vector pcDNA3. 1, or pcDNA3. 1-TROP -2, the protein levels of TROP-2 were determined by Western blotting, respectively.The migration and invasion ability was evaluated by Transwell.The E-Cadherin and N-Cadherin protein expression in PC-3 cells was examined by Western blotting. Results TROP-2 gene eukaryotic expression vector was successfully constructed. The Western blotting showed that the expression levels of TROP-2 protein were greatly increased in PC-3 cancer cells transfected with pcDNA3. 1-TROP -2.The Transwell migration assay revealed that the number of migrating cells in Con-A,Con-B, and pcDNA3. 1-TROP-2 groups was 132. 6±2. 2, 130. 8±1. 8 and 189. 6±2. 6, respectively (P< 0. 01).The Transwell invasion assay indicated that the number of migrating cells in Con-A, Con-B, and pcDNA3. 1-TROP-2 groups was 118. 16±1. 96, 117. 52±1. 85 and 166. 38±1. 65, respectively(P< 0. 01).As Compared with the Con-A group, the E-cadherin protein expression was decreased,and the N-cadherin protein expression was increased in PC-3 cancer cells transfected with pcDNA3. 1-TROP-2. Conclusion TROP-2 overexpression can promote migration and invasion of human prostate cancer cells probably promoting the epithelial-mesenchymal transition. Key words: Trophoblast cell-surface antigens 2; Prostate carcinoma; Invasion; Epithelialmesenchymal transition

Abstract 2622: TMEFF2 inhibits human prostate cancer cell migration through its G protein activating domain.

Abstract The type I transmembrane protein with epidermal growth factor and two follistatin motifs 2 (TMEFF2) is expressed mainly in brain and prostate. Expression of TMEFF2 is deregulated in a significant fraction of primary and metastatic prostate, suggesting a role in this disease. In fact, we have previously shown that TMEFF2 can function as a tumor suppressor in vitro, inhibiting monolayer and anchorage-independent cellular growth in HEK293T cells and sarcosine-induced cell migration and invasion in benign prostate epithelial RWPE-1 cells. However, the molecular mechanisms involved in the tumor suppressor phenotype of TMEFF2 are not clear. TMEFF2 has several biologically important features. The extracellular domain contains two follistatin domains and an epidermal growth factor-like (EGF) domain, while the transmembrane domain and short cytoplasmic tail have features resembling a G protein activating domain, suggesting that it may signal through a G protein. We hypothesize that the tumor suppressor activity of TMEFF2 is, at least in part, due to signaling mediated by the putative G protein activating domain. In order to test this hypothesis, here we investigate the role of TMEFF2 and a mutant lacking the cytoplasmic tail on cellular migration of prostate cancer cells, and begin analyzing the molecular mechanisms involved in this effect. Our results demonstrate that overexpression of TMEFF2 in human prostate cancer cell line RWPE-2 causes about 60% decrease in cellular migration as assessed by wound-healing assays. Interestingly, we found that RWPE-2 cells overexpressing TMEFF2 have a defect in cell spreading on culture dishes (∼20% decrease) as well as on vitronectin-coated coverslips (∼30% decrease) and that this defect is accompanied by abolished focal adhesion and stress fiber formation. Moreover, the inhibition of TMEFF2 on cellular migration is dependent on its G protein activating domain as deletion of this domain rescues migration, spreading on vitronectin, and focal adhesion formation. The specific G protein pathway(s) involved in the anti-migratory effect of TMEFF2 is currently under investigation. In summary, the data presented here indicate, for the first time, that the anti-migratory effect of TMEFF2 in human prostate cancer cells is mediated by its G protein activating domain. These results raise the possibility that TMEFF2 agonists may be used as therapeutic agents to mediate tumor suppression via this pathway. Citation Format: Xiaofei Chen, C. Justus, L Yang, M.j. Ruiz-Echevarria. TMEFF2 inhibits human prostate cancer cell migration through its G protein activating domain. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2622. doi:10.1158/1538-7445.AM2013-2622

Bisphenol A stimulates human prostate cancer cell migration via remodelling of calcium signalling

Bisphenol A (BPA), the principal constituent of reusable water bottles, metal cans, and plastic food containers, has been shown to be involved in human prostate cancer (PCa) cell proliferation. The aim of the present study was to explore the effect of BPA on PCa cell migration and the pathways involved in these processes. Using the transwell technique, we clearly show for the first time that the pre-treatment of the cells with BPA (1–10 nM) induces human PCa cell migration. Using a calcium imaging technique, we show that BPA pre-treatment induces an amplification of Store-Operated Calcium Entry (SOCE) in LNCaP cells. RT-PCR and Western blot experiments allowed the identification of the ion channel proteins which are up-regulated by BPA pre-treatments. These include the Orai1 protein, which is known as an important SOCE actor in various cell systems, including human PCa cells. Using a siRNA strategy, we observed that BPA-induced amplification of SOCE was Orai1-dependent. Interestingly, the BPA-induced PCa cell migration was suppressed when the calcium entry was impaired by the use of SOCE inhibitors (SKF96365, BTP2), or when the extracellular calcium was chelated. Taken together, the results presented here show that BPA induces PCa cells migration via a modulation of the ion channel protein expression involved in calcium entry and in cancer cell migration. The present data provide novel insights into the molecular mechanisms involved in the effects of an environmental factor on cancer cells and suggest both the necessity of preventive measures and the possibility of targeting ion channels in the treatment of PCa cell metastasis.

Notch Activation Is Dispensable for D, L-Sulforaphane-Mediated Inhibition of Human Prostate Cancer Cell Migration

D, L-Sulforaphane (SFN), a synthetic racemic analog of broccoli constituent L-sulforaphane, is a highly promising cancer chemopreventive agent with in vivo efficacy against chemically-induced as well as oncogene-driven cancer in preclinical rodent models. Cancer chemopreventive effect of SFN is characterized by G2/M phase cell cycle arrest, apoptosis induction, and inhibition of cell migration and invasion. Moreover, SFN inhibits multiple oncogenic signaling pathways often hyperactive in human cancers, including nuclear factor-κB, Akt, signal transducer and activator of transcription 3, and androgen receptor. The present study was designed to determine the role of Notch signaling, which is constitutively active in many human cancers, in anticancer effects of SFN using prostate cancer cells as a model. Exposure of human prostate cancer cells (PC-3, LNCaP, and/or LNCaP-C4-2B) to SFN as well as its naturally-occurring thio-, sulfinyl-, and sulfonyl-analogs resulted in cleavage (activation) of Notch1, Notch2, and Notch4, which was accompanied by a decrease in levels of full-length Notch forms especially at the 16- and 24-hour time points. The SFN-mediated cleavage of Notch isoforms was associated with its transcriptional activation as evidenced by RBP-Jk-, HES-1A/B- and HEY-1 luciferase reporter assays. Migration of PC-3 and LNCaP cells was decreased significantly by RNA interference of Notch1 and Notch2, but not Notch4. Furthermore, SFN-mediated inhibition of PC-3 and LNCaP cell migration was only marginally affected by knockdown of Notch1 and Notch2. Strikingly, SFN administration to Transgenic Adenocarcinoma of Mouse Prostate transgenic mice failed to increase levels of cleaved Notch1, cleaved Notch2, and HES-1 proteins in vivo in prostatic intraepithelial neoplasia, well-differentiated carcinoma or poorly-differentiated prostate cancer lesions. These results indicate that Notch activation is largely dispensable for SFN-mediated inhibition of cell migration, which should be viewed as a therapeutic advantage as Notch activation is frequent in human prostate cancers.

Similarities and differences between effects of angiotensin III and angiotensin II on human prostate cancer cell migration and proliferation

Proliferation plays a critical role in tumor growth when cell migration is essential to invasion. The effect of Ang III and Ang II was evaluated on these important processes. Changes in the migration potential of prostate cancer cells were investigated using Wound Healing Test and a Transwell Migration Chamber with a 3μm pore size. Cell proliferation was measured with a BrdU Assay and Countess Automated Cell Counter, thus determining the influence of angiotensins on hormone-dependent (LNCaP) and hormone-independent (DU-145) human prostate cancer lines. The influence of Ang III and Ang II on classic receptors may be inhibited by Losartan or PD123319. Test peptide modulation of the AT1 and AT2 receptors was examined by Western Blot and fluorescent immunocytochemistry. The results indicate that Ang III promotes the migration of both LNCaP and DU-145 lines, whereas Ang II stimulates this process only in androgen-independent cells. Both angiotensin peptides can induce prostate cancer cell proliferation in a time- and dose-dependent manner. The obtained results show that Ang III and Ang II can modify the expression of classic receptors, particularly AT2. These results suggest that the investigated peptide can modulate cell migration and proliferation in prostate cancer cells. Angiotensins probably have a greater influence on proliferation in the early-stage prostate cancer model than hormone-independent cell lines. Assume also that Ang II can enhance the migration tendency aggressive prostate cancer cells, while Ang III does so more effective in non-metastatic cells.

Stat3 Promotes Metastatic Progression of Prostate Cancer

There are currently no effective therapies for metastatic prostate cancer because the molecular mechanisms that underlie the metastatic spread of primary prostate cancer are unclear. Transcription factor Stat3 is constitutively active in malignant prostate epithelium, and its activation is associated with high histological grade and advanced cancer stage. In this work, we hypothesized that Stat3 stimulates metastatic progression of prostate cancer. We show that Stat3 is active in 77% of lymph node and 67% of bone metastases of clinical human prostate cancers. Importantly, adenoviral gene delivery of wild-type Stat3 (AdWTStat3) to DU145 human prostate cancer cells increased the number of lung metastases by 33-fold in an experimental metastasis assay compared with controls. Using various methods to inhibit Stat3, we demonstrated that Stat3 promotes human prostate cancer cell migration. Stat3 induced the formation of lamellipodia in both DU145 and PC-3 cells, further supporting the concept that Stat3 promotes a migratory phenotype of human prostate cancer cells. Moreover, Stat3 caused the rearrangement of cytoplasmic actin stress fibers and microtubules in both DU145 and PC-3 cells. Finally, inhibition of the Jak2 tyrosine kinase decreased both activation of Stat3 and prostate cancer cell motility. Collectively, these data indicate that transcription factor Stat3 is involved in metastatic behavior of human prostate cancer cells and may provide a therapeutic target to prevent metastatic spread of primary prostate cancer.

PIP3‐dependent Rac exchanger 1 (P‐Rex1) promotes prostate cancer cell migration

Activation of Rac by guanine-nucleotide exchange factors (GEFs) has been suggested to play an important role in cell migration which is critical for tumor metastasis. Here we investigated the role of P-Rex1, a novel specific GEF for Rac, in human prostate cancer cell migration. PCR and immunofluorescent staining showed that the expression of P-Rex1 was undetectable in normal prostate epithelial cells (PrEC) and less metastatic LNCaP cells but significantly increased in more metastatic prostate cancer cell lines (DU-145, PC-3-LN4 and PC-3). Our study also revealed increased P-Rex1 protein level in human prostate tumor specimens compared to the corresponding normal prostate tissues. Using an in vitro transwell migration chamber assay, we found that NIH-3T3 conditioned media (3T3-CM) can induce prostate cancer cell migration which positively correlates with expression levels of P-Rex1. Exogenously expressed P-Rex1 enhanced Gi-coupled receptor-dependent migration of PC-3-LN4 cells up to four-fold in a dose-dependent manner. Knockdown of endogenous P-Rex1 by small interfering RNA reduced the migration of PC-3 cells. Furthermore, exogenous expression of P-Rex1 in PC-3-LN4 cells induced strong Rac-dependent lamellipodia. Our results suggest that up-regulation of P-Rex1 promotes prostate cancer cell migration and contributes to the metastasis of prostate cancer. (The Nebraska State LB595 to Y. Tu.)