Prognosis prediction model for hepatocellular carcinoma based on autophagy related genes / 生物医学工程学杂志

Autophagy is a programmed cell degradation process that is involved in a variety of physiological and pathological processes including malignant tumors. Abnormal induction of autophagy plays a key role in the development of hepatocellular carcinoma (HCC). We established a prognosis prediction model for hepatocellular carcinoma based on autophagy related genes. Two hundred and four differentially expressed autophagy related genes and basic information and clinical characteristics of 377 registered hepatocellular carcinoma patients were retrieved from the cancer genome atlas database. Cox risk regression analysis was used to identify autophagy-related genes associated with survival, and a prognostic model was constructed based on this. A total of 64 differentially expressed autophagy related genes were identified in hepatocellular carcinoma patients. Five risk factors related to the prognosis of hepatocellular carcinoma patients were determined by univariate and multivariate Cox regression analysis, including TMEM74, BIRC5, SQSTM1, CAPN10 and HSPB8. Age, gender, tumor grade and stage, and risk score were included as variables in multivariate Cox regression analysis. The results showed that risk score was an independent prognostic risk factor for patients with hepatocellular carcinoma ( HR = 1.475, 95% CI = 1.280-1.699, P < 0.001). In addition, the area under the curve of the prognostic risk model was 0.739, indicating that the model had a high accuracy in predicting the prognosis of hepatocellular carcinoma. The results suggest that the new prognostic risk model for hepatocellular carcinoma, established by combining the molecular characteristics and clinical parameters of patients, can effectively predict the prognosis of patients.

Natural compounds modulate the autophagy with potential implication of stroke

Stroke is considered a leading cause of mortality and neurological disability, which puts a huge burden on individuals and the community. To date, effective therapy for stroke has been limited by its complex pathological mechanisms. Autophagy refers to an intracellular degrading process with the involvement of lysosomes. Autophagy plays a critical role in maintaining the homeostasis and survival of cells by eliminating damaged or non-essential cellular constituents. Increasing evidence support that autophagy protects neuronal cells from ischemic injury. However, under certain circumstances, autophagy activation induces cell death and aggravates ischemic brain injury. Diverse naturally derived compounds have been found to modulate autophagy and exert neuroprotection against stroke. In the present work, we have reviewed recent advances in naturally derived compounds that regulate autophagy and discussed their potential application in stroke treatment.

Construction and Validation of Prognostic Risk Score Model of Autophagy Related Genes in Lung Adenocarcinoma / 中国肺癌杂志

BACKGROUND@#Autophagy related genes (ARGs) regulate lysosomal degradation to induce autophagy, and are involved in the occurrence and development of a variety of cancers. The expression of ARGs in tumor tissues has a great prospect in predicting the survival of patients. The aim of this study was to construct a prognostic risk score model for lung adenocarcinoma (LUAD) based on ARGs.@*METHODS@#5,786 ARGs were obtained from GeneCards database. Gene expression profiles and clinical data of 395 LUAD patients were collected from The Cancer Genome Atlas (TCGA) database. All ARGs expression data were extracted, and The ARGs differentially expressed were identified by R software. Survival analysis of differentially expressed ARGs was performed to screen for ARGs with prognostic value, and functional enrichment analysis was performed. The least absolute selection operator (LASSO) regression and Cox regression model were used to construct a prognostic risk scoring model for ARGs. The receiver operating characteristic (ROC) curve was drawn to obtain the optimal cut-off value of risk score. According to the cut-off value, the patients were divided into high-risk group and low-risk group. The area under curve (AUC) and the Kaplan-Meier survival curve was plotted to evaluate the model performance, which was verified in external data sets. Finally, univariate and multivariate Cox regression analysis was applied to evaluate the independent prognostic value of the model, and its clinical relevance was analyzed.@*RESULTS@#Survival analysis, Lasso regression and Cox regression analysis were used to construct a LUAD prognostic risk score model with five ARGs (ADAM12, CAMP, DKK1, STRIP2 and TFAP2A). The survival time of patients with low-risk score in this model was significantly better than that of patients with high-risk score (P<0.001). The model showed good prediction performance for LUAD in both the training set (AUCmax=0.78) and two external validation sets (AUCmax=0.88). Risk score was significantly associated with the prognosis of LUAD patients in univariate and multivariate Cox regression analyses, suggested that risk score could be a potential independent prognostic factor for LUAD. Correlation analysis of clinical characteristic showed that high risk score was closely associated with high T stage, high tumor stage and poor prognosis.@*CONCLUSIONS@#We constructed a LUAD risk score model consisting of five ARGs, which can provide a reference for predicting the prognosis of LUAD patients, and may be used in combination with tumor node metastasis (TNM) staging for prognosis prediction of LUAD patients in the future.

Expression of miR-378 in cervical cancer and its effects on the proliferation and invasion of cancer cells as well as the underlying mechanism / 中国基层医药

Objective:To detect the expression of miR-378 in cervical cancer and investigate its effects on the proliferation and invasion of cancer cells as well as the underlying mechanism.Methods:A total of 185 cervical tissue samples of women who received gynecological examination in Qilu Hospital of Shandong University from January 2012 to January 2016 were included in this study. Reverse transcription-quantitative polymerase chain reaction was performed to determine the expression of miR-378 in cervical tissue and C-33A cells. Western blot assay was performed to detect the expression of different cancer genes ATG12, CCND1 and pRb in C-33A cells. BrdU cell proliferation and Transwell invasion assay were performed to determine cell proliferation and invasion. Target Scan was used to predict and screen miR-378 gene targets and verified by a dual-luciferase reporter assay system.Results:The expression of miR-378 in cervical intraepithelial neoplasia (CIN) III lesioned tissue and cervical cancer tissue was significantly higher than that in normal cervical tissues ( F = 103.091, t = 9.381, 8.936, both P < 0.05). The expression of miR-378 in cervical cancer tissues with positive lymph node metastasis was significantly higher than that in cervical cancer tissues with negative lymph node metastasis ( t = 1.007, P < 0.01). The overexpression of miR-378 in cervical cancer tissues significantly promoted the migration and invasion of C-33A cells ( t = 5.285, P < 0.05), while low expression of miR-378 in cervical cancer tissues significantly inhibited the migration and invasion of HeLa cells ( t = 2.941, P < 0.05). The overexpression of miR-378 in C-33A cells significantly decreased the expression of ATG12, CCND1and pRb ( t = 1.382, 1.431 and 2.086, all P < 0.05). The low expression of miR-378 in C-33A cells significantly increased the expression of ATG12, CCND1 and pRb ( t = 3.961, 3.062 and 2.894, all P < 0.05). Conclusion:miR-378 can greatly promote the metastasis of cervical cancer cells. ATG12, as a direct target of miR-378, provides new insights into the molecular mechanism underlying cervical cancer pathology and therapeutic target.

Recommendations for the use of the acetaminophen hepatotoxicity model for mechanistic studies and how to avoid common pitfalls

Acetaminophen (APAP) is a widely used analgesic and antipyretic drug, which is safe at therapeutic doses but can cause severe liver injury and even liver failure after overdoses. The mouse model of APAP hepatotoxicity recapitulates closely the human pathophysiology. As a result, this clinically relevant model is frequently used to study mechanisms of drug-induced liver injury and even more so to test potential therapeutic interventions. However, the complexity of the model requires a thorough understanding of the pathophysiology to obtain valid results and mechanistic information that is translatable to the clinic. However, many studies using this model are flawed, which jeopardizes the scientific and clinical relevance. The purpose of this review is to provide a framework of the model where mechanistically sound and clinically relevant data can be obtained. The discussion provides insight into the injury mechanisms and how to study it including the critical roles of drug metabolism, mitochondrial dysfunction, necrotic cell death, autophagy and the sterile inflammatory response. In addition, the most frequently made mistakes when using this model are discussed. Thus, considering these recommendations when studying APAP hepatotoxicity will facilitate the discovery of more clinically relevant interventions.

Expression of autophagy related gene 5 and cyclin E in coronary heart disease and its clinical significance / 中南大学学报(医学版)

OBJECTIVES@#To explore the expression of autophagy related genes 5 (ATG5) and cyclin E in coronary heart disease (CHD) and its clinical significance.@*METHODS@#From April 2018 to August 2018, 80 patients diagnosed with CHD in the Second Xiangya Hospital, Central South University were selected as an observation group, and another 80 healthy subjects were selected as a control group. The expression of ATG5 and cyclin E mRNA in nucleate cells and the plasma protein in the 2 groups were detected and analyzed. The model of macrophage-derived foam cells induced by oxidized low density lipoprotein (ox-LDL) was used to simulate atherosclerosis. The proliferation of macrophage- derived foam cells and the protein levels of ATG5 and cyclin E induced by ox-LDL at different concentrations were examined.@*RESULTS@#Compared with the control group, the levels of ATG5 mRNA and protein in the blood in the observation group were decreased, and the cyclin E mRNA and protein levels were increased, there were statistically difference (both <0.05). Receiver operating characteristic (ROC) curve showed that the area under curve (AUC) of ATG5 mRNA, cyclin E mRNA, ATG5 protein and cyclin E protein were 0.739, 0.780, 0.671 and 0.807, respectively. Pearson analysis showed that the ATG5 mRNA was negatively correlated with the cyclin E mRNA (=-0.734, <0.05),while the plasma ATG5 protein was negatively correlated with the plasma cyclin E protein (=-0.746, <0.05). Macrophage-derived foam cell model induced by ox-LDL showed that the proliferation of foam cells and the expression levels of cyclin E protein were increased in a concentration and time-dependent manner, and the expression levels of ATG5 protein were decreased in a concentration-dependent manner.@*CONCLUSIONS@#The levels of ATG5 mRNA and protein are lowly expressed while the levels of cyclin E mRNA and protein are highly expressed in the patients with CHD.The ATG5 protein levels are lowly expressed in ox-LDL-treated macrophage-derived foam cells while the cyclin E protein levels are highly expressed in ox-LDL-treated macrophage-derived foam cells. Based on these observations, we conclude that ATG5 inhibits the degradation of the cyclin E and promotes the proliferation of macrophages, involving in the occurrence and development of CHD.

Effect of Qiyu Sanlong Decoction in Inhibiting Growth of Lung Cancer by Regulating Expressions of Beclin1, Atg5 and LC3B / 中国实验方剂学杂志

Objective::To study the effect of Qiyu Sanlong decoction on the growth of subcutaneous tumor in lung cancer mice and the expressions of key autophagy molecule, yeast Atg6 homologous (Beclin1), autophagy related genes5 (Atg5), and microtubule-associated protein1 light chain3 (LC3B). Method::Lewis lung carcinoma cells (LLC) were used to reproduce the lung cancer mice transplanted model. After the modeling, the mice were randomly divided into model group, Qiyu Sanlong decoction group, chemotherapy group and combination group, with 18 transplanted mice in each group. In model group, mice were fed with 0.9% saline 20 mL·kg-1 daily. In Qiyu Sanlong decoction group, mice were fed with Qiyu Sanlong decoction 80.48 g·kg-1 daily. The chemotherapy group was intraperitoneally injected with 0.4 mL cisplatin solution (DDP) at the 1st, 3rd and 5th day. The combination group was orally given the drugs at the concentration of 80.48 g·kg-1, and 0.4 mL DDP solution was intraperitoneally injected at the 1st, 3rd and 5th day. After 21 days of continuous treatment, tumor tissue was exfoliated and weighed, and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was used to observe the histological changes of tumor. The expressions and localizations of Beclin1 and LC3B in tumor tissues were detected by immunohistochemical staining. Protein expressions of Beclin1, Atg5, LC3B-Ⅰand LC3B-Ⅱ were determined by Western blot, and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated. The transcription levels of Beclin1, Atg5 mRNA in tumor tissues were detected by Real-time PCR. Result::Qiyu Sanlong decoction had a mild inhibitory effect on transplanted tumor, with an inhibitory rate of 31.2%. Under microscope, patchy necrotic tumor cells were observed in the tumor tissues of Qiyu Sanlong decoction group. Immunohistochemical staining and Western blot analysis showed that Qiyu Sanlong decoction could up-regulate the expressions of Beclin1, Atg5 and LC3B protein (P<0.01), and promote the conversion from LC3B-Ⅰ into LC3-Ⅱ compared with the model group. Real-time PCR results showed that Qiyu Sanlong decoction could promote the transcription of Beclin1 mRNA and Atg5 mRNA compared with the model group (P<0.01). Conclusion::Qiyu Sanlong decoction has a mild inhibitory effect on lung tumors, and its mechanism may be related to up-regulating the expressions of autophagy key proteins Beclin1, Atg5 and LC3B, and promoting the conversion from LC3B-Ⅰ to LC3B-Ⅱ.

Autophagy and epigenetic modification in inflammatory bowel disease / 中国组织工程研究

BACKGROUND: Inflammatory bowel disease is a chronic inflammatory disease associated with intestinal immune, and autophagy is a cell approach to promote immune regulation. Abnormal expression of autophagy-related genes is closely related to intestinal inflammation and immune response. However, the mechanism by which epigenetic modification regulates autophagy in inflammatory bowel disease has not been fully clarified. OBJECTIVE: To introduce the role of epigenetic modification in autophagy, and to promote a further understanding of inflammatory bowel disease. METHODS: A computer-based online research of PubMed database was performed with the key words of “inflammatory bowel disease, autophagy, autophagy-related genes, epigenetic modification, DNA methylation, histone modification, chromatin remodeling, miRNA.” The search time was from January 1998 to April 2019. Finally, 61 eligible articles were included in result analysis. RESULTS AND CONCLUSION: Epigenetic modifications such as DNA methylation, histone modification, chromatin remodeling, non-coding RNA can regulate intestinal inflammation, immune and autophagy through susceptibility genes AGL and IRGM, thereby mediating the occurrence and development of inflammatory bowel disease.

Advances in autophagy-related genes in inflammatory diseases / 解放军医学杂志

Autophagy is a highly conserved physiological and biochemical process in which organisms maintain homeostasis of the cellular environment. Autophagy is widely involved in the occurrence and development of many diseases, especially in inflammatory diseases, which is a hot topic in recent years. Autophagy-related genes (ATGs) play important roles in regulating autophagy in many aspects, thus affecting the process and prognosis of inflammatory diseases. This article reviews the regulatory roles and mechanisms of autophagy in inflammatory diseases, as well as the functions and roles of several important ATGs in inflammatory diseases.

The influence and mechanism of cisplatin on autophagy and proliferation of endometrial cancer / 实用医学杂志

Objective To explore the effects of cisplatin on the proliferation and autophagy of endometrial carcinoma Ishikawa cells.Methods Ishikawa cell proliferation was detected by MTS assay after the cells were treated with CDDP.To assess the level of autophagy,transmission electron microscope and Western blot were used to detect LC3 and Beclin1 expression;fluorescence microscopy was used to observe the fluorescence aggregation of green fluorescent protein and microtubule associated protein 1 light chain 3 fusionprotein (GFP-LC3).Results Cisplatin of 10 g/mL inhibited the proliferation of Ishikawa cells,with an increase of time and concentration,the inhibition of cell proliferation was significantly elevated (P ＜ 0.01).Transmission electron microscopy showed that under a condition of cisplatin on Ishikawa endometrial cancer cell,autophagy occurred.With an increase of concentration and dosage,Western blot showed that autophagy related protein LC3 expression was up-regulated,but becline-1 had no obvious change.LC3 expression level was higher in 12h-treatment with 20 μg/mL cisplatin group than in the control group,and was higher in 24h-treatment group than in 12h-treatment group.Conclusions Cisplatin inhibits proliferation of Ishikawa cells and induces autophagy of the cells in a time-and dose-dependent manner.Autophagy related MAP-LC3 is involved in the molecular mechanisms of autophagy induced by cisplatin in endometrial carciHom.

Association of Single Nucleotide Polymorphisms of Autophagy-related Genes Atg7 Site rs11706903 and Parkin-son's Disease / 中国康复理论与实践

Objective To investigate the relationship between the single nucleotide polymorphisms(SNP)of rs11706903,intron region of Atg7,and Parkinson's disease(PD).Methods From January,2013 to March,2017,130 PD patients and 109 healthy subjects were recruit-ed and collected the blood samples.SNPs of rs11706903 were detected with polymerase chain reaction restriction fragment length polymor-phism.Results For the patients,the genotype frequencies of rs11706903 were AA 10.00%,AC 52.31% and CC 37.69%;and allele frequen-cies were A 36.15% and C 63.85%.For the healthy controls,the genotype frequencies of rs11706903 were AA 7.34%,AC 49.54% and CC 43.12%;and allele frequencies were A 32.11% and C 67.89%.There was no significant difference in frequencies of genetypes and alleles be-teen two groups(χ 2<1.001,P>0.05).Conclusion There might be no relationship between SNPs of Atg7 intron region rs11706903 and PD.

The analysis of correlation between single nucleotide polymorphism at Atg7 rs14016 and Parkinson's disease / 中华老年医学杂志

Objective To investigate the association between rs14016 (19 +31C/T) polymorphisms of autophagy-related genes 7 (Atg7) and Parkinson's disease (PD) in Han population in China.Methods Totally 123 patients with Parkinson's disease (PD) (case group) and 101 synchronized health controls (control group) were selected from Chinese Han population between January 2013 and July 2016.A single nucleotide polymorphism (SNP) of rs14016 of Atg7 gene was detected by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis.After gene sequencing for genotyping and detection of alleles,genotype and allele frequency distributions were analyzed in the two groups.Results The frequency distributions of TT genotype and T alleles were 17.9％ (22/123) and 41.1％ (101/246) in the case group,and 5.9％ (6/101) and 31.2 ％ (63/202) in the control group,respectively.The difference in genotype frequencies between the two groups was statistically significant (x2 =7.236,P =0.007,OR =3.01,95 ％ CI:1.27-7.14).The frequencies of T and C alleles were 41.1％ (101/246) and 58.9％ (145/246) respectively in the case group,and 31.2％ (63/202) and 68.8％ (139/202) in the control group (x2 =4.655,P=0.031,OR=1.32,95 ％CI:1.02-1.70),with the statistically significant difference in the allele.The TT genotype of rs14016 showed statistical significance between the two groups by logistic regression analysis (OR=3.40,95％CI:1.32-8.80,P=0.012).Conclusions The T allele and TT genotype at the rs14016 of Atg7 gene might be associated with PD,and might increase the risk for suffering from PD,which is worthy of further fully researches.