Mutation of Spastin Affects Microtubule Dynamics Through Differential Distribution
Liang Zhi,
Cai Zhenbin,
Zhang Guowei,
Tan Minghui,
Lin Hongsheng
Issue:
Volume 8, Issue 2, December 2020
Pages:
22-26
Received:
21 July 2020
Accepted:
29 July 2020
Published:
5 August 2020
Abstract: SPG4 gene encodes Spastin, and its mutations are the main cause of hereditary spastic paraplegia (HSP). There are more than 50 gene mutations in HSP, and mutations of Spastin account for more than 40%, so SPG4 is the key gene that causes the disease. In order to clarify the effect on microtubule stability after inactivation of AAA functional domain caused by mutations of Spastin, we carried out PCR amplification and C413Y site-directed mutation of the target gene according to the sequence of SPG4 in PUBMED gene bank and constructed GFP-Spastin and GFP-Spastin C413Y recombinant plasmids. The recombinant plasmids were introduced into COS7 cells, and the expression of recombinant plasmids in COS7 cells and changes of microtubule stability were observed. The results of colony PCR and gene identification showed that recombinant plasmids were successfully constructed. Western blotting showed that GFP-Spastin and GFP-Spastin C413Y could be expressed normally in COS7 cells. Immunofluorescence assay showed that the distribution of GFP-Spastin was punctate in the cells and GFP-Spastin had strong microtubule cleavage ability, while GFP-Spastin C413Y was almost distributing in the nucleus, and the ability of microtubule cleavage was weakened and the microtubule was in a stable state. Therefore, we concluded that Spastin C413Y changed the distribution of Spastin in cells and weakened the ability of microtubule cleavage. Spastin C413Y accumulated in the nucleus which could not cut microtubules effectively.
Abstract: SPG4 gene encodes Spastin, and its mutations are the main cause of hereditary spastic paraplegia (HSP). There are more than 50 gene mutations in HSP, and mutations of Spastin account for more than 40%, so SPG4 is the key gene that causes the disease. In order to clarify the effect on microtubule stability after inactivation of AAA functional domain...
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siRNA-dependent Gene Silencing of EVI1 Decreased Cell Proliferation in MKN45 Cells
Yuri Na,
Hyejun Hwang,
Jaehyuk Cho,
Woo Rin Lee
Issue:
Volume 8, Issue 2, December 2020
Pages:
27-32
Received:
14 September 2020
Accepted:
29 September 2020
Published:
16 October 2020
Abstract: MDS1 and EVI1 complex locus protein EVI1 (MECOM) is an oncogenic transcription factor in many types of cancer. However, the clinical significance of MECOM in esophagogastric cancer has not been well elucidated. The aim of this study was to define the association of amplification of MECOM and esophagogastric cancer patients’ overall survival rate and to investigate the functional role of MECOM in MKN45 cell proliferation and apoptosis. In this study, a total of 3236 esophagus and gastric patients were analyzed from 16 genomic studies using cBioPortal, which provides an open-access resource with multiple cancer genomics data sets. When the patient samples were divided into two subgroups (MECOM amplified group and MECOM non-amplified group), the poor survival rate was significantly associated with MECOM amplification (p= 0.04). To investigate the role of EVI1 on esophagogastric cancer cells, siRNA dependent gene silencing of EVI1 in MKN45 cells was conducted. RT-PCR was used to examine the expression of EVI1 in MKN45 cells. The MTT assay was used to examine cell proliferation. The apoptotic cells were quantified by fluorescence cell counter. Knockdown of EVI1 leads to reduced cell proliferation and induced apoptosis in MKN45 cells. These results indicate that MECOM may be a potential target for esophagus and gastric cancer gene-targeted therapy. Collectively, our result suggested that EVI1 is a probable target gene for esophagogastric cancer cell proliferation.
Abstract: MDS1 and EVI1 complex locus protein EVI1 (MECOM) is an oncogenic transcription factor in many types of cancer. However, the clinical significance of MECOM in esophagogastric cancer has not been well elucidated. The aim of this study was to define the association of amplification of MECOM and esophagogastric cancer patients’ overall survival rate an...
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Anti-wrinkle Effect of a Palmitoyl Oligopeptide Complex on Human Keratinocytes and Fibroblasts Through TGF-β1 Pathway
Yina Lu,
Guoxun Wang,
Dong Liu,
Jun Tian
Issue:
Volume 8, Issue 2, December 2020
Pages:
33-38
Received:
29 October 2020
Accepted:
9 November 2020
Published:
19 November 2020
Abstract: Many researchers revealed that dermal fibroblast which mainly synthesis the collagen and elastin in dermis were affects by skin ageing, such as the proliferation rate and gene expression level. Oligopeptides were verified by many reports on anti-aging focus on collagen synthesis, but few literatures revealed the molecular mechanism of oligopeptides on skin aging. The potential anti-ageing effect and the mechanism of PTC, a new palmitoyl oligopeptide complex, were evaluated. MTT assay was adopted to detect the anti-aging effect of PTC in both human dermal fibroblast HDF-a and keratinocyte NHEK. The mechanism of PTC was detected further by full transcriptome analysis using RNA-sequencing method, and ELISA analysis of key proteins. Skin wrinkles were measured by VC98 in vivo. PTC showed positive effect in cell proliferation and the biosynthesis of collagens, intergrins not only on fibroblasts, but also on keratinocytes. Moreover, many pathways were confirmed in the anti-aging effect of PTC, such as TGF-β1 pathway and ECM-receptor interaction. A cream contained 5.0% PTC showed significant improvement of skin wrinkles and roughness. So that, PTC has a potentially beneficial effect on the cell proliferation, the biosynthesis of ECM related proteins and the expression of many genes which are involved in cell cycle and secretory features to slow down skin aging. The molecular pathway maybe targets the TGF-β1 in both epidermis and dermis.
Abstract: Many researchers revealed that dermal fibroblast which mainly synthesis the collagen and elastin in dermis were affects by skin ageing, such as the proliferation rate and gene expression level. Oligopeptides were verified by many reports on anti-aging focus on collagen synthesis, but few literatures revealed the molecular mechanism of oligopeptides...
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