Effect of reinforcing qi for resolving masses method on expressions of MEK2 and p-ERK protein in estrogen induced uterine leiomyoma model guinea pigs' uterine tissue / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of reinforcing qi for resolving masses method (RQRMM) on expressions of extracellular signal regulated kinase 2 (MEK2) and phosphorylation extracellular signal regulated kinase (p-ERK) protein in estrogen induced uterine leiomyoma model Guinea pigs' uterine tissue.</p><p><b>METHODS</b>Guinea pigs were randomly divided into five groups, i.e., the model group, the high dose group, the middle dose group, the low dose group, and the Western medicine group (mifepristone). The normal control group was set up. The uterine leiomyoma model in guinea pigs was established by castrating and subcutaneous injecting estradiol (E2). The protein expression levels of MEK2 and p-ERK of guinea pigs' uterine tissues were detected by immunohistochemical assay.</p><p><b>RESULTS</b>The protein expressions of MEK2 and p-ERK in the uterine muscular tissue of Guinea pigs' uterine tissue were higher in the model group than in the normal group (P < 0.01). They decreased to some degree in the high dose group, the middle dose group, and the low dose group. Of them, the protein expressions of MEK2 and p-ERK were significantly lower in the high dose group than in the model group and the Western medicine group (P < 0.01).</p><p><b>CONCLUSION</b>RQRMM could treat uterine leiomyoma possibly through intervening the MAPK/ERK cell signal pathway to inhibit the proliferation of myoma cells.</p>

Bibenzyl from Dendrobium inhibits angiogenesis and its underlying mechanism / 药学学报

Bibenzyl is a type of active compounds abundant in Dendrobium. In the present study, we investigated the inhibitory effects of six bibenzyls isolated from Dendrobium species on vascular endothelial growth factor (VEGF)-induced tube formation in human umbilical vascular endothelial cells (HUVECs). All those bibenzyls inhibited VEGF-induced tube formation at 10 micromol x L(-1) except tristin, and of which moscatilin was found to have the strongest activity at the same concentration. The lowest effective concentration of moscatilin was 1 micromol x L(-1). Further results showed that moscatilin inhibited VEGF-induced capillary-like tube formation on HUVECs in a concentration-dependent manner. Western blotting results showed that moscatilin also inhibited VEGF-induced phosphorylation of VEGFR2 (Flk-1/KDR) and extracellular signal-regulated kinase 1/2 (ERK1/2). Further results showed that moscatilin inhibited VEGF-induced activation of c-Raf and MEK1/2, which are both upstream signals of ERK1/2. Taken together, results presented here demonstrated that moscatilin inhibited angiogenesis via blocking the activation of VEGFR2 (Flk-1/KDR) and c-Raf-MEK1/2-ERK1/2 signals.

MEK1 and MEK2 differentially regulate human insulin- and insulin glargine-induced human bladder cancer T24 cell proliferation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Increased risk of bladder cancer has been reported in diabetic patients. This study was to investigate the roles of mitogen-activated protein kinase kinase (MEK) 1 and 2 in the regulation of human insulin- and insulin glargine-induced proliferation of human bladder cancer T24 cells.</p><p><b>METHODS</b>In the absence or presence of a selective inhibitor for MEK1 (PD98059) or a specific siRNA for MEK2 (siMEK2), with or without addition of insulin or glargine, T24 cell proliferation was evaluated by cell counting kit (CCK)-8 assay. Protein expression of MEK2, phosphorylation of ERK1/2 and Akt was analyzed by Western blotting.</p><p><b>RESULTS</b>T24 cell proliferation was promoted by PD98059 at 5 - 20 µmol/L, inhibited by siMEK2 at 25 - 100 nmol/L. PD98059 and siMEK2 remarkably reduced phosphorylated ERK1/2. Insulin- and glargine-induced T24 cell proliferation was enhanced by PD98059, suppressed while not blocked by siMEK2. Insulin- and glargine-induced ERK1/2 activation was blocked by PD98059 or siMEK2 treatment, whereas activation of Akt was not affected.</p><p><b>CONCLUSION</b>MEK1 inhibits while MEK2 contributes to normal and human insulin- and insulin glargine-induced human bladder cancer T24 cell proliferation.</p>

Brain derived neurotrophic factor induces endothelial cells angiogenesis through AKT and ERK1/2 signal pathway / 中国实验血液学杂志

Our previous studies have demonstrated the effects of brain derived neurotrophic factor (BDNF) on promoting proliferation of multiple myeloma (MM) cells and inducing angiogenesis in MM in vitro. To further investigate whether the PI3K/Akt and MEK1/ERK pathway play a role in the BDNF-induced angiogenesis in vitro and to explore the further molecular mechanisms, two ways to establish human myeloma xenograft animal model were developed, their advantages and disadvantages were elucidated. The phosphorylation of AKT and ERK1/2 were detected in human umbilical vein endothelial cells (HUVECs) by Western blot. The angiogenic activity in vitro was evaluated by transwell migration assay and tubule formation assay. Cell proliferation was determined by crystal violet staining. Cell apoptosis was detected by FITC-Annexin-V/PI double staining and flow cytometry. The results showed that the BDNF activated the PI3K/Akt and MEK1/ERK pathway in the time-dependent manner. Ly294002 and PD98059 blocked the activation of Akt and ERK1/2 respond to BDNF. 100 ng/ml BDNF significantly increased HUVEC tube formation, migration and proliferation in vitro at a similar degree of 25 ng/ml VEGF. Furthermore, tube formation of HUVECs toward BDNF was significantly inhibited by 57% and 40% with 20 micromol/L Ly294002 and 20 micromol/L PD98059 treatment, respectively. At the same time, Ly294002 and PD98059 reduced the BDNF-induced migration of HUVECs by 74% and 36%, respectively. While BDNF-induced survival was only blocked by Ly294002 and BDNF-induced proliferation was only inhibited by PD98059. It is concluded that BDNF promotes angiogenesis of HUVECs in vitro. ERK and Akt are two crucial events in BDNF-mediated signal transduction leading to HUVECs angiogenesis by different mechanisms. Moreover, the latter is more important.

Parathyroid hormone 1-34 induce cardiac myocytes hypertrophy via extracellular regulated protein kinase 1/2 pathway / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the role of mitogen activated protein kinase kinase 1 (MAPKK, MEK1) and regulated kinase1/2 (ERK1/2) on cardiac hypertrophy induced by rat parathyroid hormone1-34 (rPTH1-34).</p><p><b>METHOD</b>Neonatal rat cardiomyocytes was treated with or without 10(-7) mol/L rPTH1-34 in the absence or presence 2 x 10(-5) mol/L PD98059, a MEK1 inhibitor. Cellular diameter was measured by Motic Images Advanced 3.0 software and the synthetic rate of protein in cardiac myocytes was detected by 3H-leucine incorporation, mRNA expression of atrial natriuretic peptide (ANP) was measured by RT-PCR and protein expression of ERK1/2 and p-ERK1/2 was measured by Western blot.</p><p><b>RESULTS</b>rPTH1-34 (10(-7) mol/L) significantly increase cellular diameter (+13.6 microm), 3H-leucine incorporation (+898 cpm/well), ANP mRNA expression (+73.9%), and p-ERK1/2 protein expression (+15%) compared to control cells (all P < 0.05) and these effects could be significantly attenuated by PD98059: cellular diameter (-7.1 microm), 3H-leucin e incorporation (-644 cpm/well), ANP mRNA expression (-52.2%), and protein expression of p-ERK1/2 (-18%) (all P < 0.05 vs. PTH group). PD98059 did not affect control cells without PTH treatment (all P > 0.05).</p><p><b>CONCLUSIONS</b>PD98059 attenuates PTH induced cardiac hypertrophy in vitro via inhibiting the expression of ERK1/2 and p-ERK1/2.</p>

Inhibitory effect of Hsp70 on angiotensin II-induced vascular smooth muscle cell hypertrophy

Angiotensin II (Ang II), which is an important mediator of both vascular responsiveness and growth, has been shown to induce vascular smooth muscle cell (VSMC) hypertrophy via the activation of a complex series of intracellular signaling events. Heat shock protein 70 (Hsp70) has recently been shown to protect against Ang II-induced hypertension. In this study, we tested the hypothesis that Hsp70 can protect VSMC from Ang II-induced hypertrophy. We treated VSMCs with Ang II to induce hypertrophy and to activate MAPK signaling pathway. We observed that the augmentation of Hsp70 expression inhibited Ang II-stimulated VSMC hypertrophy. This inhibitory effect of Hsp70 appears to be partly due to extracellular signal-regulated kinase (ERK1/2) inactivation, which in turn, may possibly result from the accumulation of MAP kinase phosphatase-1 (MKP-1).

Significance of MEK-ERK cascade in the development of human breast carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the MEK and ERK expression and their relationship with clinicopathological parameters in human breast carcinoma, and the effect of preoperative chemotherapy on MEK and ERK protein expression.</p><p><b>METHODS</b>Samples were obtained from 56 patients with breast carcinoma and 8 patients with benign tumors. Sixteen of the 56 patients received preoperative chemotherapy. Western blot and immunohistochemistry were used to measure the expression of MEK1, MEK2 and ERK1, ERK2 protein.</p><p><b>RESULTS</b>MEK2 and ERK1, ERK2 protein levels were increased in breast carcinoma tissue compared with those in adjacent normal tissues (t = 7.244, 5.959, 3.735, P < 0.01) and benign tumors (t = 2.206, P < 0.05). The levels of MEK1 were decreased. The expression of MEK2 protein in ER negative patients was higher than that in ER positive ones. MEK2 protein levels were lower in patients who received preoperative chemotherapy than in those who did not.</p><p><b>CONCLUSION</b>Overexpression of MEK-ERK may play an important role in the development of human breast carcinoma. MEK and ERK protein expressions are inhibited by preoperative chemotherapy.</p>