ABSTRACT
Cerebral ischemia-reperfusion injury (CIRI) is an important pathophysiological process of ischemic stroke associated with various physiological and pathological processes, including autophagy and apoptosis. In this study, we examined the role and mechanism of long noncoding RNA CAMK2D-associated transcript 2 (C2dat2) in regulating CIRI in vivo and in vitro. C2dat2 up-regulation facilitated neuronal autophagy and apoptosis induced by CIRI. Mechanistically, C2dat2 acts as a competing endogenous RNA (ceRNA) to negatively regulate miR-30d-5p expression. More specifically, miR-30d-5p targeted the 3'-untranslated region of DNA damage-inducible transcript 4 (DDIT4) and silenced its target mRNA DDIT4. Additionally, C2dat2 binding with heat shock cognate 70/heat shock protein 90 blocked RNA-induced silencing complex assembly to abolish the miR-30d-5p targeting of DDIT4 and inhibited miR-30d-5p to silence its target mRNA DDIT4. Further analysis showed that C2dat2 knockdown conspicuously inhibited the up-regulation of DDIT4 and Beclin-1 levels and LC3B II/I ratio and the down-regulation of P62 and phosphorylated mammalian target of rapamycin (mTOR)/mTOR and phosphorylated-P70S6K/P70S6K ratio in Neuro-2a cells after oxygen-glucose deprivation/reoxygenation. This study first revealed that C2dat2/miR-30d-5p/DDIT4/mTOR forms a novel signaling pathway to facilitate autophagy and apoptosis induced by CIRI, contributing to the better understanding of the mechanisms of CIRI and enriching the ceRNA hypothesis in CIRI.
Subject(s)
Ischemic Stroke/complications , MicroRNAs/metabolism , RNA, Long Noncoding/metabolism , Reperfusion Injury/genetics , Transcription Factors/genetics , Animals , Apoptosis/genetics , Autophagy/genetics , Cell Line, Tumor , Computational Biology , Datasets as Topic , Disease Models, Animal , Gene Knockdown Techniques , Gene Regulatory Networks , Humans , Ischemic Stroke/genetics , Ischemic Stroke/pathology , Mice , Phosphorylation/genetics , RNA, Long Noncoding/genetics , Reperfusion Injury/pathology , Signal Transduction/genetics , TOR Serine-Threonine Kinases/metabolism , Up-RegulationABSTRACT
OBJECTIVE@#To investigate serum levels of long non-coding RNA (lncRNA) TUSC7 in patients with esophageal squamous cell carcinoma (ESCC), its association with clinicopathological parameters and its role in promoting tumor metastasis and invasion.@*METHODS@#Serum samples were collected from 60 patients with ESCC admitted between January, 2017 and May, 2019, with 60 age- and gender-matched healthy subjects as the control group. Serum level of TUSC7 in ESCC patients and its expression in 4 ESCC cell lines was detected with RT-qPCR. The association of serum TUSC7 level with the clinicopathological features of the patients was analyzed. KYSE-30 cell models with TUSC7 overexpression or knockdown were established, and the proliferation of the cells was examined with MTT assay and their migration and invasion were assessed using wound healing and Transwell assays. Western blotting was used to detect the cellular expressions of the proteins associated with epithelial-mesenchymal transition (EMT).@*RESULTS@#The patients with ESCC had significantly lower serum TUSC7 level than the healthy control subjects ( < 0.05). The ESCC cell lines also expressed lower levels of TUSC7 than normal cells ( < 0.05). Serum TUSC7 level was negatively correlated with tumor staging, lymph node metastasis and infiltration ( < 0.05) but was not significantly correlated with other clinicopathological parameters in ESCC patients. In the cell experiment, overexpression of TUSC7 in KYSE-30 cells significantly inhibited cell migration and invasion ( < 0.05), enhanced the expression of the EMT marker protein E-cadherin and lowered the expressions of N-cadherin, Vimentin and MMP9 ( < 0.05); knocking down TUSC7 in the cells produced the opposite effects.@*CONCLUSIONS@#The down-regulation of TUSC7 expression in the serum of ESCC patients and in ESCC cell lines is associated with the metastasis of ESCC and promotes tumor cell migration and invasion by promoting EMT, indicating the potential of serum TUSC7 level as a molecular marker for diagnosis, treatment and metastasis monitoring of ESCC.
Subject(s)
Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Epithelial-Mesenchymal Transition , Esophageal Neoplasms , Genetics , Esophageal Squamous Cell Carcinoma , Genetics , Gene Expression Regulation, Neoplastic , Neoplasm Invasiveness , RNA, Long Noncoding , GeneticsABSTRACT
Objective To evaluate the expression of WWOX gene in breast cancer and its relation to hypermethylation of WWOX gene CpG island. Methods The expression of WWOX protein was evaluated by immunohistochemical staining in breast cancer cell line and breast cancer tissues.Methylation specific PCR(MSP)was used to check whether it Was methylated in the promoter and exon 1 of WWOX.Results Deletion in the WWOX expression was observed more frequently in invasive breast tumors,(32.2%)than normal breasttissues(5.4%)(P＜0.01).20.3% of premenopause eases were completely negative for WWOX expression compared to 57.2% of postmenopause breast carcinomas(P＜0.01).Furthermore,23.1% of Stage Ⅰ(6/26),28.6%of Stage Ⅱ(10/35),46.2%of Stage Ⅲ(12/26)tunlors showed negative WWOX protein expression.In breast cancer specimens.methylation rate of promoter and exon 1 CpG island of WWOX gene was 55%and 45%respectively.Whereas.WWOX CpG islands of normal mammary tissues were completely unmethylated in all cases.CpG islands of WWOX promoter and exon 1 were methylated in MDA-MB-231 cell line but not MCF-7 cell.Conclusions Some deletion in WWOX expression is common in breast cancer and methylation of WWOX DNA CpG islands plays a crucial role in the silence of WWOX.WWOX may play a role in the carcinogenesis and development of breast cancer.
ABSTRACT
OBJECTIVE: To discuss the responsibility transformation of pharmacists and its necessity.METHODS: The process of transformation in fundamental responsibility of pharmacists were reviewed; the current situation in the development of pharmacist group was presented, and the necessity to transform the responsibility of pharmacists was analyzed.RESULTS&CONCLUSION: It is the contemporary demand to transform the responsibility of pharmacists in China.Training of pharmacy talents and the development in the field of pharmacy, construction of information system, the development and utilization of information, scientific research, the development and utilization of new technology and method, as well as the strengthened management of drug distribution chain, and the establishment of Pharmacist Law can all help to achieve a full range of pharmaceutical care.
