Effects of Regulation of SHH/Gli1 Signaling Pathway by Baishile Capsules on Hippocampal Neurogenesis Depression Model Rats / 中国中医药信息杂志

Objective To explore the effects and mechanism of Baishile Capsules regulating SHH/Gli1 signaling pathway on hippocampal neurogenesis of depression model rats.Methods Totally 32 SD rats were randomly divided into control group,model group,fluoxetine(5.4 mg/kg)group and Baishile Capsules(2.88 g/kg)group,with 8 rats in each group.A depression rat model was established using chronic unpredictable mild stress and single cage feeding method.The model was established and administered simultaneously for 21 consecutive days.Depression-like behavior in rats were evaluated by sucrose preference experiment and open field experiment,ELISA was used to detect brain derived neurotrophic factor(BDNF)contents in rat serum and hippocampal tissue,the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus was observed by immunofluorescence,immunofluorescence and Western blot were used to detect the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue.Results Compared with the control group,the degree of sucrose preference significantly decreased in the model group(P<0.01),the number of horizontal and vertical movements significantly decreased(P<0.01),the contents of BDNF in serum and hippocampal tissue significantly decreased(P<0.05),the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly decreased(P<0.01),and the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly decreased(P<0.01,P<0.05).Compared with the model group,the degree of sucrose preference and the number of horizontal and vertical movements in fluoxetine group and Baishile Capsule group increased significantly(P<0.05,P<0.01),the contents of BDNF in serum and hippocampal tissue significantly increased(P<0.05,P<0.01),and the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.05),the fluorescence intensity and protein expressions of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly increased(P<0.01,P<0.05).Conclusion Baishile Capsule can promote the hippocampus neurogenesis in depression model rats by regulating SHH/Gli1 signaling pathway,and play an antidepressant role.

Effects of pristimerin on the proliferation, apoptosis and vasculogenic mimicry of cervical cancer HeLa cells by regulating the Shh/Gli1 signaling pathway / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨扁蒴藤素（Pris）调节Shh/Gli1信号通路对宫颈癌HeLa细胞增殖、凋亡和血管生成拟态（VM）的影响及其机制。方法：采用MTT法检测不同浓度Pris对宫颈癌HeLa细胞增殖的抑制作用，以选取合适的干预浓度。将HeLa细胞分为对照组、环巴胺组、Pris组、Pris+pc-NC组和Pris+pc-Shh组。采用MTT法、EdU法检测各组细胞的增殖能力，Transwell小室法、流式细胞术检测各组细胞的迁移及侵袭能力和细胞凋亡率，体外血管生成实验观察VM形成情况，qPCR法检测各组细胞中Shh和Gli1 mRNA表达水平，WB法检测细胞中血管内皮生长因子A（VEGF-A）、血管内皮钙黏素（VE-cadherin）、Ki-67、caspase-3及与Shh/Gli1信号通路相关蛋白表达水平。结果：0.25~2.5 mmol/L的Pris对HeLa细胞增殖均有显著抑制作用，选择1.5 mmol/L的Pris进行后续实验。对照组细胞形成良好的管腔结构，与对照组相比，环巴胺组、Pris组和Pris+pc-NC组HeLa细胞管腔结构被明显破坏，细胞增殖活力和增殖率、迁移及侵袭细胞数目、Shh和Gli1 mRNA、VEGF-A、VE-cadherin、Ki-67、Shh、Gli1蛋白表达均显著降低（均P<0.05），细胞凋亡率和caspase-3表达均显著升高（均P<0.05）；环巴胺组与Pris组HeLa细胞各项检测指标比较差异均无统计学意义（均P>0.05）；与Pris+pc-NC组相比，Pris+pc-Shh组细胞管腔结构形成明显改善，细胞增殖活力和增殖率、迁移及侵袭细胞数、Shh和Gli1 mRNA、VEGF-A、VE-cadherin、Ki-67、Shh、Gli1蛋白表达均显著升高（均P<0.05），细胞凋亡率和caspase-3表达均显著降低（均P<0.05）。结论：Pris抑制宫颈癌HeLa细胞的增殖、迁移与侵袭和VM的形成并促进细胞凋亡，可能与阻断Shh/Gli1信号通路有关。

Genetic diagnosis of congenital hypopituitarism in Turkish patients by a target gene panel: novel pathogenic variants in GHRHR, GLI2, LHX4 and POU1F1 genes

ABSTRACT Objective: Congenital hypopituitarism (CH) is a rare disease characterized by one or more hormone deficiencies of the pituitary gland. To date, many genes have been associated with CH. In this study, we identified the allelic variant spectrum of 11 causative genes in Turkish patients with CH. Materials and methods: This study included 47 patients [21 girls (44.6%) and 26 boys (55.4%)] from 45 families. To identify the genetic etiology, we screened 11 candidate genes associated with CH using next-generation sequencing. To confirm and detect the status of the specific familial variant in relatives, Sanger sequencing was also performed. Results: We identified 12 possible pathogenic variants in GHRHR, GH1, GLI2, PROP-1, POU1F1, and LHX4 in 11 patients (23.4%), of which six were novel variants: two in GHRHR, two in POU1F1, one in GLI2, and one in LHX4. In all patients, these variants were most frequently found in GLI2, followed by PROP-1 and GHRHR. Conclusion: Genetic causes were determined in only 23.4% of all patients with CH and 63% of molecularly diagnosed patients (7/11) from consanguineous families. Despite advances in genetics, we were unable to identify the genetic etiology of most patients with CH, suggesting the effect of unknown genes or environmental factors. More genetic studies are necessary to understand the etiology of CH.

Comparaison de deux équations de référence pour le diagnostic des anomalies spirométriques à Cotonou / NA

Introduction : L'interprétation de la spirométrie se base sur des équations de référence, prenant en compte l'âge, le sexe, la taille et la race. Au CNHU-PPC, deux équations ont été utilisées ces dernières années : celles de l'American Thoracic Society/European Respiratory Society "ATS/ERS" (publiées en 1983, actualisées en 1993), et celles du Global Lung Initiative (GLI) 2012, désormais recommandées par plusieurs sociétés savantes. Notre étude visait à comparer les interprétations de spirométries basées sur ces équations. Matériel et méthodes : Nous avons mené une étude rétrospective descriptive, avec recrutement exhaustif des spirométries réalisées au CNHU-PPC du 1er Janvier 2018 au 31 Mars 2020. Les données recueillies furent analysées avec le logiciel R. Le coefficient Kappa a été calculé pour apprécier la performance des équations ATS/ERS par rapport au GLI 2012. Pour tous les tests statistiques, la différence était statistiquement significative pour une p-value inférieure à 0,05. Résultats : Les 955 spirométries recensées concernaient une population majoritairement féminine (sex-ratio=0,7) et jeune (âge moyen=44±20 ans). Il y avait plus de spirométries normales selon les équations ATS/ERS (53,6%, versus 53,0% selon GLI 2012 ; Kappa=0,71). Un TVO était objectivé dans 18,6% des cas selon l'ATS/ERS (versus 18,0% selon GLI 2012, Kappa=0,90). Il y avait moins de TVR selon l'ATS/ERS (21,3%, versus 29 % selon GLI 2012 ; Kappa=0,72), et moins de TVM selon l'ATS/ERS (5,3%, versus 6,6% selon GLI 2012 ; Kappa=0,79). Les proportions d'asthmatiques étaient identiques (12,3%). Les équations ATS/ERS ont objectivé moins de BPCO et de maladies restrictives (respectivement 4,6% et 21,3%) que le GLI 2012 (respectivement 5,8% avec Kappa=0,74, et 29% avec Kappa =0,72). Conclusion : Les équations ATS/ERS objectivent moins d'anomalies spirométriques que celles du GLI 2012 au CNHU-PPC. Des études ultérieures s'imposent pour intégrer les valeurs de référence béninoises au GLI 2012, actuellement récommandées pour l'interprétation de la spirométrie

Introduction: Spirometry's interpretation is based on reference equations, taking into account age, sex, height and race. At the CNHU-PPC, two equations have been used in recent years: those of the American Thoracic Society / European Respiratory Society "ATS/ERS"(published in 1983, updated in 1993), and more recently, those of the Global Lung Initiative (GLI) 2012, now recommended by several learned societies. Objectives: Our study aimed to compare interpretations of spirometry based on these equations. Material and methods: We carried out a descriptive retrospective study, with exhaustive recruitment of the spirometry done at the CNHU-PPC from January 1, 2018, to March 31, 2020. The data collected were analyzed with the software R. The Kappa coefficient was calculated to assess the performance of the ATS/ERS equations compared to GLI 2012. Results: The 955 spirometries recorded concerned a predominantly female (sex ratio=0.7) and young (mean age=44±20 years) population. There was more normal spirometry according to the ATS/ERS (53.6%, vs 53.0% according to GLI 2012; Kappa=0.71). An obstructive ventilatory disorder was objectified in 18.6% of cases according to ATS/ERS (vs 18.0% for GLI 2012, Kappa=0.90). There was less restrictive ventilation disorder according to ATS/ERS (21.3%, vs 29% for GLI 2012; Kappa=0.72), and less mixed ventilatory disorder according to ATS/ERS (5.3%, vs 6.6% for GLI 2012; Kappa=0.79). The proportions of asthma patients were identical (12.3%). ATS/ERS objectified less COPD and restrictive diseases (respectively 4.6% and 21.3%) than GLI 2012 (respectively 5.8% with Kappa=0.74, and 29% with Kappa=0.72).

DAPAGLIFLOZIN ASSOCIATED EUGLYCEMIC DIABETIC KETOACIDOSIS: A CASE REPORT

SGLT2 inhibitors are a new class of drugs for lowering blood sugar levels in type 2 diabetics. They have been shown to reduce cardiovascular risk along with improving glycemic control. Some of the SGLT2 inhibitors are Canagli?ozin, Dapagli?ozin, Empagli?ozin, Ertugli?ozin, Remogli?ozin. We are presenting a case of a 60-yearold female patient who is a known case of Type 2 Diabetes Mellitus presented to the emergency room with loss of responsiveness for 1 day gradual in onset. Her history revealed she is type 2 diabetic for the past 10yrs and was hospitalized 20days back when her RBS was 889mg/dl & urine ketones were positive with a diagnosis of type 2 DM with DKA. since then, she was put on Tab Dapagli?ozin 10mg OD along with other OHA's. On presentation, the patient was unconscious GCS-E1, V2, M2-5/15, pulse3 100/min, BP-80mm of hg systolic, glucometer RBS-211 mg/dl, ABG showed severe metabolic acidosis pH-6.86, HCO -2.9mmol/L, 2 PCO -24mm hg, PaO2-58mm hg, urine ketones came positive, and the patient was managed conservatively. The patient responded well, and her GCS improved with stabilization in her condition. Dapagli?ozin and other SGLT2 inhibitors can cause Euglycemic DKA, and these can be missed out in the emergency room as they have not so high blood sugar levels making the diagnosis of DKA dif?cult in emergency conditions

The role of nuclear transcription factor Gli in the regulation of hepatic epithelial mesenchymal transition in biliary atresia mice caused by Rhesus monkey rotavirus infection / 中华实用儿科临床杂志

Objective:To investigate the role of nuclear transcription factor Gli1/Gli2 of the sonic hedgehog (Shh) signaling pathway in the hepatic epithelial mesenchymal transition (EMT) of biliary atresia mice caused by Rhesus rotavirus (RRV) infection.Methods:The biliary atresia model in mice was generated by RRV infection.Mice were divided into normal group, model group, Gli1 overexpression group, Gli1 shRNA group, Gli2 overexpression group and Gli2 shRNA group.Real-time fluorescence quantitative polymerase chain reaction and Western blot were used to detect the mRNA and protein expressions of regulatory factors for EMT (Snail/Slug) and characteristic cytokines of EMT [Vimentin, α-smooth muscle actin(α-SMA), E-cadherin] in mouse liver tissues.Additionally, hematoxylin-eosin staining and Masson staining were performed to calculate the percentage of liver fibrous tissue expression area.The data were analyzed by One- Way ANOVA and LSD- t test. Results:The relative mRNA expression of Snail, Slug, Vimentin, α-SMA and E-cadherin in Gli2 overexpression group, Gli2 shRNA group and model group were 15.13±3.40, 5.48±0.46, 8.78±1.06, 12.40±2.18 and 3.06±0.53; 3.73±1.16, 5.62±1.75, 3.56±1.06, 3.88±1.16 and 10.51±1.83; 8.13±1.27, 5.32±0.98, 5.05±0.98, 4.02±0.77 and 5.12±1.60.Compared with those of the model group, mRNA levels of Snail, Vimentin and α-SMA were significantly higher in Gli2 overexpression group, while that of E-cadherin was significantly lower( t=4.53, 5.29, 8.12, -2.13; all P<0.05); compared with those of the model group, mRNA levels of Snail and Vimentin in Gli2 shRNA group significantly decreased, while that of E-cadherin significantly increased( t=-2.86, -2.12, 5.62; all P<0.05). In Gli2 overexpression group, Gli2 shRNA group and model group, the protein levels of Snail, Slug, Vimentin, α-SMA and E-cadherin were 2.02±0.39, 0.31±0.08, 0.95±0.17, 1.07±0.17 and 0.42±0.06; 0.53±0.13, 0.40±0.18, 0.20±0.04, 0.28±0.07 and 1.09±0.31; 0.70±0.15, 0.42±0.22, 0.64±0.13, 0.81±0.11 and 0.42±0.09.Compared with those of the model group, protein levels of Snail, Vimentin and α-SMA were significantly higher in Gli2 overexpression group( t=12.71, 4.28, 3.70; all P<0.05); compared with those of the model group, protein levels of Vimentin and α-SMA in Gli2 shRNA group significantly decreased, while that of E-cadherin significantly increased( t=-6.14, -7.57, 5.96; all P<0.05). However, no significant change trend were detected in expression levels of characteristic cytokines of EMT between Gli1 overexpression group and Gli1 shRNA group.The area percentage of liver fiber expression in normal group, model group, Gli1 overexpression group, Gli1 shRNA group, Gli2 overexpression group and Gli2 shRNA group were (1.03±0.58)%, (33.02±11.39)%, (39.81±5.67)%, (26.06±1.29)%, (49.81±8.57)% and (17.55±0.66)%, respectively.Besides, in terms of percentage of area expressed in liver fiber tissue, the Gli2 overexpression group and Gli2 shRNA group were statistically significant compared with the model group( t=3.21, -2.96; all P<0.05), while the Gli1 overexpression group and Gli1 shRNA group were not statistically significant compared with the model group (all P>0.05). Conclusions:The Shh signaling pathway plays an important role in liver fibrosis in mice with biliary atresia.Gli2, a key transcription factor of Shh signaling pathway, can significantly regulate liver EMT process in mice with biliary atresi.

Expression and significance of GLI1 and Shh in the malignant transformation of ovarian endometriosis / 中华妇产科杂志

Objective:To investigate the expression levels and clinical significance of glioma-associated oncogene homolog 1 (GLI1) and sonic hedgehog signaling molecule (Shh) in the malignant transformation of ovarian endometriosis (EM).Methods:The expressions of GLI1 and Shh were detected by real-time reverse transcription (RT)-polymerase chain reaction (PCR) and EnVision method in 50 cases of ovarian EM tissues, 35 cases of atypical endometriosis (aEM) and 50 cases of endometriosis-associated ovarian cancer (EAOC). The expression differences of two molecular markers in the malignant transformation of ovarian EM were compared, and the relationships between two molecular markers and the clinicopathological features and prognosis of EAOC were analyzed.Results:(1) RT-PCR showed that the expression levels of GLI1 mRNA in EM, aEM and EAOC group were 1.77±0.40, 3.54±0.44, and 7.80±0.24, respectively. The expression levels of Shh mRNA were 0.95±0.21, 3.14±0.35, and 5.41±0.31, respectively. GLI1 and Shh mRNA in EAOC group were significantly higher than those in EM and aEM group (all P<0.01), and there were statistically significant differences between EM and aEM group (all P<0.01). The percentages of GLI1 in ovarian EM, aEM and EAOC were 32% (16/50), 57% (20/35), and 66% (33/50), respectively, meanwhile, the positive expression rates of Shh were 20% (10/50), 49% (17/35), and 54% (27/50), respectively (all P<0.01). GLI1 mRNA expression was positively correlated with Shh mRNA expression in EAOC tissues ( r=0.721, P<0.01). The expressions of GLI1 protein were proportionated to Shh protein in EAOC tissues ( r=0.608, P=0.001). (2) The expression of GLI1 was significantly related to the International Federation of Gynecology and Obstetrics (FIGO) stage, cancer antigen 125 (CA 125) levels, lymph node metastasis, and Platinum resistance in EAOC patients (all P<0.05). The expression of Shh were related to FIGO stage and lymph node metastasis in EAOC patients (all P<0.05). Logistic regression analysis showed that GLI1 expression was an independent risk factor for poor prognosis in EAOC patients ( P<0.05). Kaplan-meier survival analysis showed that the overall survival rate of EAOC patients with high GLI1 expression and low GLI1 expression was 12.1% and 35.3%, respectively, with statistical significance ( χ2=10.73, P<0.01). The overall survival rate of EAOC patients with high and low expression of Shh protein was 11.1% and 30.4%, in which there was statistically significant difference ( χ2=3.96, P=0.047). Conclusion:GLI1 and Shh are highly associated with the malignant transformation of ovarian EM, which may play a role in promoting malignant degeneration of ovarian EM, and the high expression of GLI1 and Shh indicates a poor prognosis in EAOC patients.

Mechanism of Cordycepin Inhibiting Proliferation and Promoting Apoptosis of Hepatoma Cells by Regulating Gli1 / 中国实验方剂学杂志

ObjectiveTo investigate the molecular mechanism of cordycepin inhibiting proliferation and promoting apoptosis of human hepatoma cells （HCCs）. MethodGlioma-associated oncogene homolog 1 （Gli1） gene was silenced by small interfering RNA （siRNA） and transfected into SMMC-7721 cells， and then cell proliferation was detected by cell counting kit-8 （CCK-8） assay and cell cloning assay. SMMC-7721 cells were treated with different concentration of cordycepin， and the cell proliferation and apoptosis were examined. The expression of Gli1 and the downstream related genes was determined by Real-time polymerase chain reaction（PCR） and Western blot. ResultThe mRNA and protein expression of Gli1 in SMMC-7721 cells was higher than that in normal liver cells （P<0.01）. The proliferation rate of SMMC-7721 with silenced Gli1 decreased at 72 and 96 h （P<0.05， P<0.01）， and the colony-forming capacity lowered （P<0.01） compared with those in the blank group. Compared with the control， 80 μmol·L-1 and 120 μmol·L-1 cordycepin significantly inhibited the proliferation of SMMC-7721 cells at 72 and 96 h （P<0.05， P<0.01）， and promoted the apoptosis of them （P<0.01）. Moreover， 80 and 120 μmol·L-1 cordycepin restrained the mRNA and protein expression of Gli1 in SMMC7721 cells （P<0.05， P<0.01）. At 120 μmol·L-1， cordycepin led to the decrease in the mRNA and protein levels of B-cell lymphoma-2 （Bcl-2） and c-Myc （P<0.05， P<0.01）， and the increase in the mRNA and protein expression of cysteine-aspartic acid protease-3 （Caspase-3） （P<0.05）. ConclusionGli1 is highly expressed in HCCs， and cordycepin can suppress the proliferation and enhance the apoptosis of HCCs by regulating Gli1 and the downstream apoptosis-related factors.

Glioma del nervio óptico en paciente adulto con presentación hemorrágica tratado mediante abordaje endoscópico transesfenoidal extendido / Optic nerve glioma in an adult patient with hemorrhagic presentation treated by an extended transsphenoidal endoscopic approach

El glioma del nervio óptico es una entidad de muy baja incidencia en pacientes adultos, lo cual impide tener suficiente información sobre historia natural y conducta terapéutica en este grupo etario. En el presente artículo comunicamos el caso de un paciente de 27 años de edad con compromiso agudo del nervio óptico izquierdo debido a hemorragia intra tumoral, forma de presentación muy poco común en este tipo de tumores. Se realizó la resección mediante un abordaje endoscópico transesfenoidal extendido, con preservación funcional de la vía óptica contralateral. La anatomía patológica confirmó astrocitoma pilocítico positivo para el rearreglo KIAA 1549-BRAF. y negativo para la mutación BRAF V600E. Teniendo en cuenta la histopatología y biología molecular en este caso, la estabilidad visual contralateral y la resección quirúrgica amplia, se decidió no realizar tratamiento adyuvante con radioterapia o quimioterapia. El objetivo de esta conducta fue evitar lesiones adicionales sobre el quiasma, nervio óptico contralateral y/o hipotálamo. Dada la escasa información existente en la literatura médica, el reporte de este caso podría contribuir con información adicional en el manejo y conducta terapéutica de este tipo de lesiones.

The optic nerve glioma is a very uncommon entity in adult patients, with little information about its natural history and therapeutical management. We report the case of a 27-year-old patient with acute involvement of the left optic nerve due to intratumoral hemorrhage, a very uncommon form of presentation in this type of tumor. Resection was performed using an extended transsphenoidal endoscopic approach, with functional preservation of the contralateral optic pathway. The histopathology confirmed positive pilocytic astrocytoma with KIAA 1549-BRAF rearrangement and without BRAF V600E mutation. Considering the histopathology and molecular biology, the contralateral visual stability and the wide surgical resection, it was decided not to perform further treatment. The purpose of this decision was to avoid additional damage to the chiasm, contralateral optic nerve and/or hypothalamus. Given the limited data available in medical literature, the report of this case could contribute with additional information on the management and therapeutic approach of this type of tumors

The role of Hedgehog signaling pathway in the simvastatin-induced osteogenic differentiation of bone marrow mesenchymal stem cells / 国际药学研究杂志

Objective: To investigate the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSC)induced by simvastatin(SIM)in vitro and then further investigate whether the Hedgehog signaling pathway is involved in the SIM-induced osteogenic differentiation of BMSC using the Hedgehog signaling pathway blocking agent cyclopamine(Cpn). Methods: The rat BMSC was extracted by the whole-bone marrow adherent culture method and cul- tured in the induction medium(control medium,CM)and the induction medium containing SIM or SIM+Cpn. The expression of alkaline phosphatase(ALP)in induced cells was detected by the ALP staining. Immunofluorescence stain- ing was performed to detect the espressions of Gli1 and osteocalcin(OCN)in the induced cells. The Gli1,ALP,colla- gen type(COL)and OCN mRNA expression was detected by real-time quantitative PCR(RT-PCR). Western blot (WB)was used for assessing the expression level of Gli1,Runx2,COLand OCN proteins. The cell calcium nodule for- mation and matrix mineralization ability were detected by alizarin red staining. Results: The ALP expression was signifi- cantly higher in SIM group than in CM group(P<0.05),while the ALP expression was lower in SIM+Cpn group than in the SIM group(P<0.05)but higher than in the CM group. Immunofluorescence assay showed that SIM could promote the expression of Gli1 and OCN in the conditions with or without Cpn. On the 10th and 18th day of intervention,the mRNA expression level of Gli1,ALP,OCN and COLwas significantly higher in the SIM group than in the CM group(P<0.05), and the higher mRNA expression in the SIM group could be completely blocked by Cpn. Further,on the 10th and 18th day of intervention,the expressed Gli1,Runx2,COLand OCN protein level was significantly higher in the SIM group in the CM group(P<0.05),while the level of these expressed proteins in the SIM+Cpn group was significantly lower than in the SIM group(P<0.05). The alizarin red staining showed that SIM group had stronger matrix mineralization ability than the other groups(P<0.05). Conclusion: SIM could up-regulate the expression of Gli1 and the osteogenesis markers ALP,Runx2,COLand OCN to induce osteogenic differentiation of BMSC,and Cpn could not completely block the SIM-induced differentiation. Further study is needed to uncover the underlying mechanism.

Research progress on zinc finger protein GLi in development and treatment of hepatocellular carcinoma / 国际外科学杂志

Hepatocellular carcinoma is one of the most common malignant tumors in the world,and its morbidity and mortality have been high.The current preferred treatment for hepatocellular carcinoma is surgical treatment and liver transplantation,but the 5-year survival rate is still low.Due to the insidious onset of hepatocellular carcinoma,most patients have no special manifestations at the initial stage.At the time of diagnosis,they are already in the terminal stage and lose the opportunity of surgery.The Hh pathway plays a key role in the development of tumors.The zinc finger protein GLi plays an important role in tumorigenesis as a key transcription factor in the Hh pathway.At the same time,zinc finger protein GLi can play a key role in the invasion and metastasis of hepatocellular carcinoma by inducing epithelial-mesenchymal transition.In addition,the zinc finger protein GLi also interacts with various tumor-related factors to affect the occurrence and development and treatment of hepatocellular carcinoma.This article reviews the role of zinc finger protein GLi in the development and treatment of hepatocellular carcinoma.

Anti-pancreatic fibrosis of oxymatrine by promoting expression of transcription factors Gli2 in LTC-14 and PANC-1 pancreatic stellate cells / 中草药

Objective: To investigate the effect of oxymatrine (OM) on the expression of Gli2 in LTC-14 cells and PANC-1 cells induced by TGF-β1. Methods: LTC-14 cells and PANC-1 cells were divided into four groups:control group, TGF-β1 group, TGF-β1 + OM group, and OM group. The protein was extracted after 12 h, and the expressions of related proteins were detected by Western blot. Results: Compared with control group, the expression of Gli2 in LTC-14 cells was significantly decreased induced by TGF-β1, while the expressions of α-SMA, FN, and CoL-I were significantly increased. The expressions of Gli2, FN, CoL-I, and α-SMA were increased significantly induced by TGF-β1when compared with those of control group in PANC-1 cells, and the expression of Gli2 was inhibited by OM pretreatment in TGF-β1 + OM group compared with TGF-β1 group. OM pretreatment can increase the expression of Gli2 before stimulating with TGF-β1 in TGF-β1 + OM group in comparison with TGF-β1 group. Conclusion: OM plays an protected role in pancreatic fibrosis through promoting the expression of Gli2 in LTC-14 cells.

Effect of up-regulation of Shh-Gli1 signaling pathway on radioresistance and cell cycle in human esophageal cancer cell line Eca109 / 中国病理生理杂志

AIM:To investigate the relationship between Sonic Hedgehog(Shh)signaling pathway and cell cycle and radioresistance of esophageal cancer by up-regulating Gli1,a key factor in Shh signaling pathway.METHODS:The human esophageal cancer cell line Eca 109 was transfected with plasmid to induce Gli 1 over-expression,which served as Eca109-ox-Gli1 group.In addition, Eca109 cells transfected with empty plasmid served as negative control group and the untreated Eca109 cells were used as normal control group.The expression of Gli1 was confirmed by real-time PCR and Western blot.The radiosensitivity of the cells in the 3 groups was determined by colony formation assay.The effect of irra-diation on the cell cycle was analyzed by flow cytometry.RESULTS:The expression of Gli1 in Eca109-ox-Gli1 group was higher than that in the other 2 groups(P<0.05).The survival fraction at dose of 2 Gy in Eca109-ox-Gli1 group was high-er than that in normal control group, indicating that the radioresistance of the Eca 109 cells transfected with Gli1 plasmid was increased.The cells in Eca109-ox-Gli1 group showed higher S phase proportion than that in normal control group and negative control group(P<0.01).After irradiation at dose of 6 Gy,all cells in the 3 groups found that the cell cycle was arrested at G2/M phase,while the cells in normal control group showed higher G 2/M phase proportion than that in Eca109-ox-Gli1 group(P<0.01).CONCLUSION: The up-regulation of Gli1 may enhance the radioresistance of esophageal cancer by regulating the cell cycle.

Effect of Gli2 on apoptosis and ROS level in renal tubular epithelial cells under high glucose / 中国免疫学杂志

Objective:To investigate the effect of Gli2 on apoptosis and ROS level in renal tubular epithelial cells under high glucose. Methods:The renal tubular epithelial cell NRK-52E was used as the object of study,Gli2 overexpression vector and control vector were transfected,the non transfected cells were also used as controls,RT-PCR and Western blot detected Gli2 levels,culture of non transfected cells by high glucose and low glucose cell culture medium, cells transfected with Gli2 overexpression vector were cultured in high glucose medium,apoptosis was detected by flow cytometry,the ROS content was detected by DCFH-DA probe method, the content of SOD was detected by xanthine oxidase method; Bax, Cleaved Caspase-3 and Smo levels were detected by Western blot. Results:The Gli2 mRNA and protein levels of NRK-52E cells transfected with Gli2 overexpression vector were significantly higher than that of control cells(P＜0. 01),the Gli2 mRNA and protein levels in the NRK-52E cells transfected with the control vector were not different from those of the control cells(P＞0. 05). The apoptosis rate of NRK-52E cells without transfection was elevated after high glucose culture,elevated ROS content,SOD content decreased,the levels of Bax,Cleaved Caspase-3 were elevated in the cells,Smo levels declined,compared with the control cells,the difference was statistically significant(P＜0. 01). After overexpression of Gli2 over-expression vector,NRK-52E cells were cultured with high glucose,the rate of apoptosis decreased,ROS content decreased,elevated SOD content,the levels of Bax,Cleaved Caspase-3 decreased,elevated levels of Smo,compared with NRK-52E cells cultured with simple high glucose,the difference was statistically significant(P＜0. 01). Conclusion:Apoptosis and oxidative damage in renal tubular epithelial cells induced by high glucose, Gli2 can decrease the apoptosis of renal tubular epithelial cells induced by high glucose, mitigate oxidative damage.

Crosstalk of Hedgehog/Gli and PI3k/Akt Pathway Promotes Epithelial-mesenchymal Transition in Gastric Adenocarcinoma / 中国医科大学学报

Objective To investigate the tumor molecular mechanism of Hedgehog/Gli in promoting the epithelial-mesenchymal transition (EMT) in gastric cancer AZ521 cells. Methods After 24 h of treatment with GANT61,the mRNA expression of Gli1,Gli2, N-cadherin,and E-cadherin in the AZ521 cell line were detected by real-time fluorescence quantitative PCR. A Western blotting assay was conducted to determine the expression of the above cytokines,p-AKT and AKT. The effect of GANT61 on invasion was observed by transwell assay. N-Shh stimulation of the Hedgehog pathway was conducted to confirm the changes in these cytokines. Results GANT61 significantly downregulated the mRNA expression of Gli1,Gli2,and N-cadherin,but upregulated E-cadherin mRNA expression. The Western blotting assay revealed that GANT61 downregulated the protein expression of Gli1,Gli2,p-AKT,and N-cadherin,but upregulated E-cadherin expression. Furthermore,GANT61 inhibited the invasion. N-Shh proteins up-regulated Gli1,Gli2,and N-cadherin mRNA,protein expression and p-AKT protein expression,but downregulated E-cadherin mRNA and protein expressions. N-Shh promoted the invasion of tumor cells. Conclusion Downregulation of Gli1 and Gli2 can inhibit the invasion and metastasis in gastric cancer cells,which may be related to the promotion of EMT by Gli through the PI3K/AKT pathway.

miR-7 inhibits activation of astrocytes derived from rats via silencing of EGFR / 中国药理学通报

Aim To explore the effects of miR-7 on astrocyte activation and the underlying mechanisms. Methods Following isolation and culturing, astro-cytes extracted from rat cortex were treated with culture solution (control group), ciliary neurotrophic factor (CNTF, an agonist of astrocyte activation), miR-7 mimic+CNTF, miR-7 mimic control+CNTF, miR-7 inhibitor+CNTF and miR-7 inhibitor control+CNTF, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA ex-pression of glial fibrillary acidic protein (GFAP) and epidermal growth factor receptor(EGFR). The protein expression of GFAP, EGFR, signal transducers and activators of transcription 3(STAT3) and phosphoryla-ted STAT3 (p-STAT3) was measured using Western blot. Wild type pGL3-EGFR and mutant pGL3-EGFR-m recombinant plasmids were constructed and then co-transfected with miR-7 mimic into HEK293T cells,re-spectively. The luciferase activity of reporter gene was measured. In addition,astrocytes were treated with ei-ther EGFR siRNA or S31-201 (an inhibitor of STAT3),followed by the incubation with miR-7 inhib-itor and CNTF. Both qRT-PCR and Western blot were subsequently used to detect the mRNA and protein lev-els of GFAP. Results The expression levels of GFAP and EGFR as well as p-STAT3/STAT3 ratio in CNTF group were higher than those in control group (P <0.01). When compared with CNTF group,GFAP and EGFR levels and p-STAT3/STAT3 ratio significantly decreased in miR-7 mimic+CNTF group but increased in miR-7 inhibitor+CNTF group(P<0.01). In com-parison with control group, transfection with miR-7 mimic markedly reduced the luciferase activity of wild type EGFR (P <0.01). Moreover, miR-7 inhibitor-induced up-regulation of GFAP expression was almost completely reversed by either EGFR siRNA or S31-201 pretreatment (P<0.01). Conclusion miR-7 antag-onizes the activation of astrocytes from rats by inhibi-ting the EGFR/STAT3 signaling pathway.

Identification of Diverse Adenosine-to-Inosine RNA Editing Subtypes in Colorectal Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: RNA editing generates protein diversity by altering RNA sequences in coding regions without changing the overall DNA sequence. Adenosine-to-inosine (A-to-I) RNA editing events have recently been reported in some types of cancer, but they are rare in human colorectal cancer (CRC). Therefore, this study was conducted to identify diverse RNA editing in CRC. MATERIALS AND METHODS: We compared transcriptome data of 39 CRC samples and paired adjacent tissues from The Cancer Genome Atlas database to identify RNA editing patterns in CRC, focusing on canonical A-to-I RNA edits in coding sequence regions. We investigated nonsynonymous RNA editing patterns by comparing tumor and normal tissue transcriptome data. RESULTS: The number of RNA edits varied from 12 to 42 per sample. We also observed that hypoand hyper-RNA editing patterns were distinguishable within the samples. We found 10 recurrent nonsynonymous RNA editing candidates in nine genes (PDLIM, NEIL1, SRP9, GLI1, APMAP, IGFBP7, ZNF358, COPA, and ZNF587B) and validated some by Sanger sequencing and the inosine chemical erasing assay. We further showed that editing at these positions was performed by the adenosine deaminase acting on RNA 1 enzyme. Most of these genes are hypoedited in CRC, but editing of GLI1 was increased in cancer tissues compared with normal tissues. CONCLUSION: Our results show that nonsynonymous RNA editing patterns can be used to identify CRC patients and could serve as novel biomarkers for CRC.

The research progress of Gli1 in the regulation of epithelial mesenchymal transition in oral squamous cell car-cinoma / 口腔疾病防治

@#HH (hedgehog) signal pathway consists the hedgehoge ligand (Shh、Ihh and Dhh), the Patch and Smo membrane protein complex and Zinc finger transcription factor Gli (Gli1, Gli2, Gli3). In HH (hedgehog) signaling pathway, Gli1 not only plays a dominant and decisive role in the zinc finger transcription factor Gli (Glioma-associated oncogene homolog) family, but also includes epithelial-mesenchymal transition (EMT) and oral squamous cell carcinoma, tumor invasion and metastasis. The research on oral squamous cell carcinoma and Shh/Gli1 signal axis is rare. In this paper, the squamous cell carcinoma of oral epithelial mesenchymal transformation and correlation study of quality Gli1 is reviewed.

Effects of oxymatrine on expression of related molecules in Smad3/Gli1 signaling pathway in PANC-1 cells induced by TGF-β1 / 中草药

Objective: To investigate the effect of oxymatrine (OM) on expression of related molecules in Smad3, Gli1 signaling pathway in PANC-1 cells induced by TGF-β1. Methods: TGF-β1-induced PANC-1 cells were used to establish the pancreatic fibrosis model in vitro, and observe the effects of OM pretreatment on the related molecular expression of Smad3/Gli1. Gli1 and Smad3 RNA interference plasmids were transfected into PANC-1 cells. The protein expression levels of Smad3, Gli1 and α-SMA were measured by Western blotting. The levels of fibronectin (FN) and type I collagen (CoL-I) in the supernatant of cell culture were detected by ELISA. Results: Compared with the control group, the protein expressions of Smad3, Gli1 and α-SMA increased significantly in PANC-1 cells after treated with TGF-β1. The expressions of Gli1, α-SMA, FN, and CoL-I in PANC-1 cells decreased significantly after Gli1 RNA interference plasmid transfection compared with TGF-β1 induced group. The expression of Smad3, Gli1, α-SMA, FN, and CoL-I also decreased significantly in PANC-1 cells after Smad3 RNA interference plasmid transfection compared with TGF-β1 group. Conclusion: OM could prevent pancreatic fibrosis by regulating TGF-β1/Smad3/Gli1 signaling pathway in PANC-1 cells.

Expression of Gli-1 and EMT-related proteins and its clinical significance in non-small cell lung cancer / 中国肿瘤临床

Objective:To investigate the expression and clinical significance of glioma cancer-related gene homologous protein 1 (Gli-1) and epithelial-mesenchymal transition (EMT)-related proteins, namely, Snail, E-cadherin, and N-cadherin in non-small cell lung cancer (NSCLC). Methods:Immunohistochemical staining was performed to determine the expression levels of Gli-1, Snail, E-cadherin, and N-cadherin in 67 cases of NSCLC and 20 cases of normal lung tissues and its relationship with clinicopathological features and prognosis was explored. Another 20 samples of fresh NSCLC tissues and corresponding normal lung tissues were collected to detect the mRNA level through reverse transcription polymerase chain reaction (RT-PCR). Results:1) The positive rates of Gli-1, Snail, E-cadherin, and N-cadherin in NSCLC were 61.19％(41/67), 50.75％(34/67), 56.72％(38/67), and 53.73％(36/67), respectively;whereas those of normal lung tissues were 20％(4/20), 10％(2/20), 100％(20/20), and 5％(1/20), respectively. These two sets of data have significant statistical difference (P<0.05). 2) The high expression of Gli-1 in tumor tissues was closely related to lymph node metastasis, tumor, node, and metastasis (TNM) stage, and tumor differentiation (P<0.05) but was not associated with gender, age, tumor size, and pathological type. The expression of Gli-1 in NSCLC tissues was negatively correlated with E-cadherin (r=-0.325, P<0.05) and positively correlated with Snail and N-cadherin (r=0.379, r=0.490, P<0.05). 3) RT-PCR results showed that the expression levels of Gli-1, Snail, and N-cad-herin mRNA were significantly higher in NSCLC cases than in normal lung cases (P<0.05). The expression level of E-cadherin mRNA was lower in tumor tissues than in lung tissues (P<0.05). 4) Patients with high expression levels of Gli-1, Snail, and N-cadherin had signifi-cantly worse prognosis and lower survival rate than those with low expression (all P<0.05), whereas patients with low expression lev-els of E-cadherin had significantly better prognosis and lower survival rate than those with high expression (P<0.05). Multivariate Cox's proportional hazard regression analysis indicated that E-cadherin-negative group expression, lymph node metastasis, TNM stage, and tumor differentiation were independent prognostic factors for NSCLC. Conclusion: The abnormal activation of Hedgehog signaling pathway in NSCLC is correlated with EMT. Detecting the expression levels of Gli-1 and EMT-related proteins Snail, E-cadherin, and N-cadherin might be helpful in understanding the clinicopathological features and prognosis of patients with NSCLC.