Tellers Tale: The Journals of Esoph


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Introduction

Cover artwork courtesy of N. Alexandra Blake. Visit her blog at: aretroremedy. Home Synopsis Contact Blog. Teller's Tale Some speak of the moon. Contact Contact the author at or submit this form. Next, we performed a dual luciferase reporter assay to assess its transcriptional activity. It was reported that phosphorylation of ERK, which could be induced by EGF stimulation or gain-function mutation of Kras, was critical for ELK1-mediated ternary complex formation and transactivation [ 24 , 25 ].

So we added EGF to the culture medium to promote phosphorylation of ELK1 and noticed that the luciferase ratio was dramatically increased in dose-dependent manner Figure 5 G. SLP2 promotes proliferation and colony formation rate of GC cell in vitro and tumor growth in vivo. C group were transfected with Cas9 lentivirus and then non-target sgRNA lentivirus. D , indicated cells were plated in 6 well culture-plates for colony formation.

C: non-target sgRNA transfected , and tumor growth was monitored after the indicated times. F , tumors derived from hind limbs of NCG mice 50 days after subcutaneous injection of indicated cells.

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G , tumor weight was determined 50 days after transplantation. Nevertheless, we constructed a plasmid that tagged triple flag label to SLP2 and transfected it into AGS cell line to distinguish exogenous and endogenous expression of SLP2. These results indicate critical role of SLP2 for the activation of MAPK signaling pathway and motivate us to explore potential reagent to control GC progression by targeting this pathway.

Sorafenib, a Raf1 inhibitor which has been administrated in several types of carcinoma [ 26 , 27 ], was added into the medium of SLP2 overexpressed AGS cell. Afterwards, we investigated the anti-proliferative effects of Sorafenib on MGC cell and found that proliferation and colony formation rate of MGC cell was significantly suppressed in vitro Figure S6 A and B.

Thus, Sorafenib might be an alternative strategy for the management of GC. However, its biological function and regulation mechanism was still undisclosed that propelled us to explore the role of SLP2 in GC especially when we noticed the growing evidence of its correlation with clinical outcome in several types of malignancy. Evidence so far has illustrated that cell-cycle dysregulation will result in uncontrolled cell proliferation and promote tumor development by initiating unscheduled cell division [ 28 ].

We reveal that deletion of SLP2 drastically suppresses cell proliferation in cultured cells and tumor growth in xenograft implants. Moreover, we found that Sorafenib, which specifically inhibits phosphorylation of Raf1, could attenuate accelerated proliferation rate by SLP2 up-regulation in GC cell. PHB was originally thought to be a tumor suppressor that played a central role in the inhibition of cell-cycle progression through interacting with p53 and retinoblastoma tumor suppressor protein RB in the nucleus [ 29 , 30 ]. Further evidences showed the anti-tumorigenic effect of PHB in prostate cancer [ 31 ] and liver cancer [ 32 ].

Despite PHB was reported with anti-tumorigenic properties, there were mounting evidence indicating its pro-tumorigenic role. PHB expression was increased in tumor tissues of the cervix [ 34 ], esophagus [ 35 ], breast [ 36 ], lung [ 37 ], bladder [ 38 ], thyroid [ 39 ], ovary [ 40 ], and prostrate [ 41 ]. Down-regulation of PHB expression drastically reduced the rate of cell division and capability to exhibit anchorage-independent growth in certain cancer cell lines [ 42 ] and PHB was shown to be necessary for the activation of Raf1 by the oncogene Ras in HeLa cells [ 22 ].

Furthermore, three independent researches revealed that GC tissues also showed higher expression compared to paired normal tissue [ 20 , 43 , 44 ]. In our study, we found that loss of PHB inhibited cell proliferation and tumor growth and provided unprecedented evidence between PHB level and prognosis of GC patients.

Reassessing the paradoxical reports of PHB, we notice that PHB is identified to be a tumor suppressor especially if the level of PHB is increased in the nucleus which might interact with p53 and RB [ 29 , 30 ], whereas elevated PHB in cytoplasm and cytomembrane, documented in many reports, may facilitate tumorigenesis [ 34 - 41 ]. Further studies are needed to clarify the distinct function of intracellular sub-localization of PHB during the progression of cancer.

SLP2 not only contributed to the stability of PHB but also retained PHB in cytoplasm that might be the most important factor to induce the dysfunction of this loop. Surprisingly, we discovered that transcription factor ELK-1, which could activate multiple oncogenes, was capable to bind to the promoter of SLP2. A , B and C , Total cell lysates prepared from the indicated cells were subjected to immunoblot with different antibodies. D , indicated cells were plated in 96 culture-plates and OD values were measured. E , indicated cells were plated in 6 well culture-plates for colony formation.

C , sequence logo for ELK1 binding derived from Consite database. Enrichment of c-fos promoter was set as a positive control for ELK1 binding. G , indicated vectors were co-transfected into T cells and cells were stimulated with or without EGF for 48 hr and cell lysates were subjected to luciferase assay. K, indicated vectors and siRNAs were co-transfected into T cells and cell lysates were subjected to luciferase assay. Sorafenib abrogates the accelerated cell proliferation induced by SLP2 elevation through inhibiting activation of MAPK signaling pathway.

D, indicated cells were plated in 96 culture-plates and OD values were measured. E, indicated cells were plated in 6 well culture-plates for colony formation. Our current study represents the first comprehensive investigation into the functional relationship and regulation mechanism between SLP2 and GC. This provides a novel mechanism of tumor promoting role of SLP2-PHB complex and disrupting their connection may be an attractive target for future therapeutic strategies in GC management.


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Furthermore, sorafenib induced inhibition of Raf1 phosphorylation similarly suppressed cell proliferation in SLP2 overexpressed cells. The correlation between expression of SLP2 and clinical pathological features in external cohort. Global cancer statistics, CA Cancer J Clin. J Clin Oncol. Cancer statistics in China, Cancer Genome Atlas Research N. Comprehensive molecular characterization of gastric adenocarcinoma. Increased levels of SLP-2 correlate with poor prognosis in gastric cancer. Gastric Cancer. Wang Y, Morrow JS. J Biol Chem. Comparative proteomics of human endothelial cell caveolae and rafts using two-dimensional gel electrophoresis and mass spectrometry.

Identification of a novel mitochondrial complex containing mitofusin 2 and stomatin-like protein 2. Stomatin-like protein 2 binds cardiolipin and regulates mitochondrial biogenesis and function. Mol Cell Biol. Extensive temporally regulated reorganization of the lipid raft proteome following T-cell antigen receptor triggering.

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