1.Expression Level and Clinical Significance of Gli1 in Patients with Myelodysplastic Syndrome.
Zhen YIN ; Yu-Ting QIN ; Chun-Yang DEN ; Tuerxun NILUPAR ; Shuang CHEN ; Huan WANG ; Yasen HALIDA ; Ming JIANG ; Jian-Ping HAO
Journal of Experimental Hematology 2019;27(3):867-871
OBJECTIVE:
To study the expression level and clinical significance of Gli1 gene in patients with myelodysplastic syndrome(MDS).
METHODS:
The positive rate of bone marrow CD34 cells was detected by flow cytometry in 53 patients with MDS.Magnetic beads were used to separate CD34 cells. The expression of Gli1 on CD34 cells was detected by RT-qPCR, 25 patients with iron deficiency anemia were selected as controls. The relationship of Gli1 expression with clinical characteristics were analyzed.
RESULTS:
The expression of Gli1 in patients with MDS (0.73±1.26) was significantly higher than that in the control group (0.07±0.46) (P<0.05). The expression of Gli1 significantly correlated with platelet count, chromosome grouping and IPSS risk stratification (P<0.05). The median overall survival time of patients in high and low expression groups were 7 and 20 months respectively (P<0.05). Multivariate analysis showed that Gli1 and chromosome grouping were 2 independent poor prognostic factors (P<0.05).
CONCLUSION
The expression of Gli1 is high in MDS. Abnormal expression of Gli1 positively correlates with clinical characteristics and prognosis of patients.Gli1 may be involved in the occurrence and development of MDS.
Bone Marrow Cells
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Flow Cytometry
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Humans
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Myelodysplastic Syndromes
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Prognosis
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Zinc Finger Protein GLI1
2.Divergent chondro/osteogenic transduction laws of fibrocartilage stem cell drive temporomandibular joint osteoarthritis in growing mice.
Ruiye BI ; Qianli LI ; Haohan LI ; Peng WANG ; Han FANG ; Xianni YANG ; Yiru WANG ; Yi HOU ; Binbin YING ; Songsong ZHU
International Journal of Oral Science 2023;15(1):36-36
The anterior disc displacement (ADD) leads to temporomandibular joint osteoarthritis (TMJOA) and mandibular growth retardation in adolescents. To investigate the potential functional role of fibrocartilage stem cells (FCSCs) during the process, a surgical ADD-TMJOA mouse model was established. From 1 week after model generation, ADD mice exhibited aggravated mandibular growth retardation with osteoarthritis (OA)-like joint cartilage degeneration, manifesting with impaired chondrogenic differentiation and loss of subchondral bone homeostasis. Lineage tracing using Gli1-CreER+; Tmfl/-mice and Sox9-CreER+;Tmfl/-mice showed that ADD interfered with the chondrogenic capacity of Gli1+ FCSCs as well as osteogenic differentiation of Sox9+ lineage, mainly in the middle zone of TMJ cartilage. Then, a surgically induced disc reposition (DR) mouse model was generated. The inhibited FCSCs capacity was significantly alleviated by DR treatment in ADD mice. And both the ADD mice and adolescent ADD patients had significantly relieved OA phenotype and improved condylar growth after DR treatment. In conclusion, ADD-TMJOA leads to impaired chondrogenic progenitor capacity and osteogenesis differentiation of FCSCs lineage, resulting in cartilage degeneration and loss of subchondral bone homeostasis, finally causing TMJ growth retardation. DR at an early stage could significantly alleviate cartilage degeneration and restore TMJ cartilage growth potential.
Animals
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Mice
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Osteogenesis
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Zinc Finger Protein GLI1
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Fibrocartilage
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Temporomandibular Joint
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Disease Models, Animal
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Osteoarthritis
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Stem Cells
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Growth Disorders
3.Expression and clinical significances of hedgehog signaling pathway in non-Hodgkin's lymphoma.
Xi-Yuan ZHANG ; Xiao-Qian LIU ; Xue-Ling GE ; Li-Li FEN ; Xin WANG
Journal of Experimental Hematology 2011;19(5):1129-1133
This study was aimed to investigate the expression and clinicopathologic significance of Gli1 and Gli2, 2 factors of Hedgehog(Hh) signaling pathway, in non-Hodgkin's lymphoma (NHL). Gli1 and Gli2 mRNA and protein in 18 cases of NHL and 10 cases of reactive lymphadenitis were amplified and identified by real-time PCR, and were assayed by immunohistochemical staining respectively. The results showed that (1) Gli1 and Gli2 mRNA in NHL group (RQ 2.05, 2.31) were expressed higher than that in reactive lymphadenitis group (RQ 0.82, 0.89). Gli1 mRNA activated level was positively related with Gli2 (r = 0.63, p < 0.01). In addition, Gli2 also positively correlated to clinical stages of NHL (p = 0.03), but the expressions of Gli1 and Gli2 mRNA had no significant correlation to B symptoms, blood β(2)-microglobulin, age and sex. (2) The positive expression rate of Gli1 and Gli2 protein in NHL group were 80% and 68% respectively, which were extremely higher than that in reactive lymphadenitis group. Gli1 protein level was positively related with Gli2 (r = 0.62, p < 0.05). Both Gli1 and Gli2 protein expression positively correlated to clinical staging of NHL (p = 0.05, p = 0.01). It is concluded that the Gli1 and Gli2 of Hh signaling pathway have been found to higher express in patients with NHL, and have significance for clinical staging and predicting prognosis of NHL. To further investigate the role of Hh signaling pathway in NHL will contribute to elucidate the occurrence and development of NHL, and provide a favorable method for therapy of NHL.
Adult
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Aged
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Female
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Hedgehog Proteins
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metabolism
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Humans
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Kruppel-Like Transcription Factors
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metabolism
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Lymphoma, Non-Hodgkin
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metabolism
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pathology
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Male
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Middle Aged
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Neoplasm Staging
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Nuclear Proteins
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metabolism
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Signal Transduction
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Transcription Factors
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metabolism
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Zinc Finger Protein GLI1
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Zinc Finger Protein Gli2
4.Effect of Gli1 gene silencing on proliferation of K562 cells and its mechanisms.
Wen-xia SU ; Yong-hong CHEN ; Wen ZENG ; Wen-li LIU ; Han-ying SUN
Chinese Journal of Hematology 2012;33(7):570-573
OBJECTIVETo investigate the effect of Gli1 gene silencing by RNA interference (RNAi) on proliferation of K562 cells and its mechanisms.
METHODSThe small interference RNA (siRNA) was synthesized in vitro. K562 cells were transfected with Gli1 siRNA by the way of lipofection (lipofectamine 2000). Non-specific siRNA transfected cells were used as control. Transfection efficiencies of different siRNA concentrations were detected by flow cytometry and the best siRNA concentration was selected. The silencing effect of siRNA was demonstrated by real time PCR and Westem blot analysis. Cell proliferation was measured by MTT method, cell cycle by PI assay, c-myc and p21 mRNA level was detected by real time PCR analysis.
RESULTSTransfection efficiency of siRNA was increased in a dose-dependent manner when siRNA concentration was below 200 pmol, and the highest transfection efficiency reached (80.11 ± 5.63)%. Both the mRNA and protein level of Gli1 was down-regulated in Gli1 specific siRNA group, the mRNA level was (52.60 ± 3.57)% of that of control group after 24 h (t = 20.33, P < 0.01) and the protein level was (79.31 ± 5.58)% of that of control group after 48 h (t = 6.54, P < 0.01). The cell proliferation rate in Gli1 siRNA group was (94.41 ± 3.58)% (t = 2.40, P = 0.05) and (90.22 ± 3.34)% (t = 4.37, P < 0.01) of that of control group after 24 h and 48 h, respectively. G(2)/M cell cycle arrest was observed, the mRNA level of c-myc was down-regulated while p21 was up-regulated in Gli1 siRNA group after 24 h and 48 h (P < 0.05).
CONCLUSIONSTargeted silencing of Gli1 gene by RNAi inhibits the proliferation of K562 cells, which acts through the down-regulation of c-myc and up-regulation of p21 expression.
Cell Proliferation ; Gene Silencing ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transcription Factors ; genetics ; Transfection ; Zinc Finger Protein GLI1
5.Expression of Gli1 in Adult Acute Lymphoblastic Leukemia and Its Clinical Significance.
Bing LONG ; Xiang-Zhong ZHANG ; Xu-Dong LI ; Zi-Jie LONG ; Xiao-Zhen WANG ; Jia-Jun LIU ; Dong-Jun LIN
Journal of Experimental Hematology 2015;23(4):946-949
OBJECTIVETo explore the expression and clinical significance of Hedgehog signaling transcription factor Gli1 in acute lymphoblastic leukemia (ALL) patients.
METHODSThe clinical specimens were obtained from 32 newly diagnosed and 6 relapsed ALL patients. Normal bone marrow cells from 15 healthy donors were used as controls. Real-time qPCR and Western blot were applied to detect Gli1 mRNA and protein expression in bone marrow mononuclear cells (BMMNC) of these samples respectively. The relation of Gli1 mRNA levels with clinical parameter was also evaluated.
RESULTSThe expression level of Gli1 mRNA in de novo and relapsed ALL patients was significantly higher than that in the normal controls (P < 0.05). There was no stalistically significant difference of the Gli1 mRNA expression between de novo and relapsed ALL cases (P > 0.05). In 24 de novo ALL patients with complete remission (CR) after induction chemotherapy, the levels of Gli1 mRNA were significantly reduced as compared with levels before treatment (P < 0.05). However, in 4 ALL patients without remission, no obvious difference of Gli1 mRNA levels were observed as compared with levels of Gli1 before treatment (P > 0.05). A positive correlation between the Gli1 mRNA expression level and white blood cell count (WBC) was found in the BMMNC of ALL patients (R = 0.725, P < 0.05). Similarly, Gli1 protein expression was significantly higher in the de novo and relapsed ALL cases compared with normal controls. The Gli1 protein level was down-regulated when the ALL patients was in CR.
CONCLUSIONThe expression of Gli1 mRNA and protein has been found to be high in de novo and relapsed ALL patients, and the change of Gli1 expression maybe relate to therapeutic efficacy and prognosis of ALL patients.
Bone Marrow Cells ; Humans ; Induction Chemotherapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Remission Induction ; Transcription Factors ; Zinc Finger Protein GLI1
6.GLI-1 is involved in EGF-regulated enhancement of the invasiveness of prostate cancer ARCaP(E) cells in vitro.
Guo-dong ZHU ; Jian-cheng ZHOU ; Jin ZENG ; Zhen-kun MA ; Bo-xing SU ; Xin-yang WANG ; Da-lin HE
National Journal of Andrology 2012;18(1):16-22
OBJECTIVETo investigate the role of the hedgehog (HH) signaling pathway transcription factor glioma-associated oncogene hoinolog 1 (GLI-1) in EGF-regulated enhancement of the invasiveness of the prostate cancer ARCaP(E) cell line in vitro.
METHODSThe expressions of EGFR and GLI-1 in prostate cancer ARCaP(E) cells were analyzed by immunofluorescence staining. ARCaP(E) cells were treated with EGF at 100 ng/ml, followed by detection of the changes in cell morphology and invasiveness, as well as in the expressions of p-ERK, ERK and GLI-1. Migration transwell assay was used to determine the effects of 100 ng/ml EGF and GLI-1 antagonist GANT61 on the invasiveness of the ARCaP(E) cells.
RESULTSBoth EGFR and GLI-1 were expressed in the ARCaP(E) cells. EGF induced morphological transition of epithelial-like ARCaP(E) cells to mesenchymal-like cells, increased their in vitro invasiveness, and significantly upregulated the expressions of p-ERK and GLI-1 in the ARCaP(E) cells (P<0.05). GANT61 significantly inhibited the in vitro invasiveness of the ARCaP(E) cells and reduced the enhancing effect of EGF on their invasiveness (P<0.05).
CONCLUSIONThe results from ARCaP(E) cells shed light on the cross-talk of the HH pathway with the EGF/ERK signaling pathway. GLI-1 might be responsible for EGF-regulated enhancement of the invasiveness of ARCaP(E) cells in vitro.
Cell Line, Tumor ; Epidermal Growth Factor ; metabolism ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; Zinc Finger Protein GLI1
7.NCTD Retards AML HL60 Cell Proliferation via Targeting Hedgehog/SOX2 axis.
Ming-Yan FU ; Wei-Wei CHEN ; Na GAO ; Shuo LI ; Jing DU ; Wen-Zheng YU
Journal of Experimental Hematology 2021;29(1):32-37
OBJECTIVE:
To investigate the effect of norcantharidin (NCTD) to proliferation of leukemia cells through disrupting key regulators of sonic Hedgehog (SHH) pathway and its downstream transcription factor SOX2.
METHODS:
CCK8 was used to detected the HL60 and NB4 cells after inhibited by NCTD, SMO and GLI1 inhibitor for 24 hours. Expression level of SMO, GLI1 and SOX2 in HL60 cells with NCTD treatment was detected by immunoblot. HL60 cells were transfected with pcDNA3.1 plasmid expressing GLI1 or SOX2. Empty vector and pcDNA3. 1-EGFP were divided into negative and positive control group, respectively. The expression of exogenous GLI1 or SOX2 in HL60 cells was confirmed by immunoblot, and growth curve of HL60 cell was checked by CCK8. Proliferation of genetic modified HL60 cells treated by various dose of NCTD was detected.
RESULTS:
NCTD, SMO/GLI1 inhibitors could inhibit the proliferation of NB4 and HL60 cells in a dose-dependent manner. Compared with solvent (DMSO)-treated control group, NCTD remarkably decreased protein level of SMO, GLI1 and SOX2. GLI1 and SOX2 were overexpressed in HL60 cells as compared with pcDNA3.1 empty vector-transfected group. Growth curve demonstrated significant proliferative advantage of GLI1/SOX2-transfected cells. CCK8 assay indicated that GLI1/SOX2-overexpressed HL60 cells were more resistant to NCTD treatment.
CONCLUSION
NCTD attenuates HL60 proliferation via targeting the Hedgehog/SOX2 axis.
Bridged Bicyclo Compounds, Heterocyclic
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Cell Proliferation
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HL-60 Cells
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Hedgehog Proteins
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Humans
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Leukemia, Myeloid, Acute
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SOXB1 Transcription Factors
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Zinc Finger Protein GLI1
8.Expression of Smo protein and the downstream transcription factor Gli1 protein in Sonic hedgehog signal transduction pathway in gastric carcinoma.
Zhen-xiang RONG ; Chi-hua FANG ; Da-jian ZHU ; Sheng-jun LIU
Journal of Southern Medical University 2006;26(12):1728-1731
OBJECTIVETo study the expression of Smo protein and the downstream transcription factor Gli1 protein in Sonic hedgehog signal transduction pathway in gastric carcinoma.
METHODSA tissue microarray was constructed using 85 gastric carcinoma and 25 normal gastric mucosa specimens. The expression of Smo and Gli1 proteins were detected immunohistochemically and the correlation between their expression in gastric carcinoma was analyzed.
RESULTSOnly weak expression, if any, of Smo and Gli1 proteins was detected in normal gastric mucosa, but in papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, their expressions were significant increased as the differentiation degree was lowered. Smo protein expression in gastric carcinoma was significantly correlated with that of Gli1 protein with correlation coefficient of 0.989 (P<0.001).
CONCLUSIONThe abnormal activity of Sonic hedgehog signal transduction pathway may play an important role in the occurrence of papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, and this abnormality is associated with Smo protein overexpression, which upregulates the expression of the downstream transcription factor Gli1 protein.
Adenocarcinoma ; metabolism ; pathology ; physiopathology ; Adult ; Aged ; Female ; Hedgehog Proteins ; physiology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Receptors, G-Protein-Coupled ; biosynthesis ; Signal Transduction ; Smoothened Receptor ; Stomach Neoplasms ; metabolism ; pathology ; physiopathology ; Transcription Factors ; biosynthesis ; Zinc Finger Protein GLI1
9.Expression patterns of sonic hedgehog signaling molecules in human fetal prostate development.
Guo-Dong ZHU ; Da-Lin HE ; Hui HE ; Lin-Lin ZHANG ; Xin-Yang WANG ; E Haiyen ZHAU ; Leland W K CHUNG
National Journal of Andrology 2006;12(10):896-899
OBJECTIVETo investigate whether the sonic hedgehog signaling pathway is involved in the development of human fetal prostate, and to evaluate the changing staining patterns of its molecules, sonic hedgehog (SHH), patchedl (PTC1), smoothened (SMO), and GLI1, in the human fetal prostate at various gestation stages.
METHODSFifteen human fetal prostate specimens at various developmental stages (10 - 39 weeks) were included in this study. SHH, PTC1, SMO and GLI1 were detected in all the specimens by immunohistochemical technique. All the slides were observed and assessed under the light microscope.
RESULTSSHH, PTC1, SMO and GLI1 could be detected in human fetal prostate tissues, and their expression formed two surges, the former at week 16, and the latter at week 28. The staining of SHH and SMO was distributed only in the ductal epithelium but not in the stroma. The expression of PTC1 and GLI1 could be found mainly in the epithelium, with minimal staining in the stroma.
CONCLUSIONThe sonic hedgehog signaling pathway is involved in the development of the human fetal prostate. The high expression of its molecules at early gestation stages might be associated with the induction of prostatic buds, while their abundant expression at later gestation stages might be related to the prostate ductal branching, growth, differentiation and morphogenesis.
Gene Expression Regulation, Developmental ; physiology ; Hedgehog Proteins ; biosynthesis ; Humans ; Male ; Oncogene Proteins ; biosynthesis ; Patched Receptors ; Prostate ; embryology ; metabolism ; Receptors, Cell Surface ; biosynthesis ; Receptors, G-Protein-Coupled ; biosynthesis ; Signal Transduction ; physiology ; Smoothened Receptor ; Trans-Activators ; biosynthesis ; Zinc Finger Protein GLI1
10.Inhibitory effect of cyclopamine on metastatic ability of EC109 cells and its mechanism.
Xiaoping ZUO ; Zhiming QIN ; Kaibin WANG ; Xiangru ZHENG ; Liqian CHEN
Journal of Southern Medical University 2012;32(12):1828-1832
OBJECTIVETo investigate the effect of cyclopamine on metastatic ability of human esophageal cancer EC109 cells and explore the possible mechanism.
METHODSTranswell chamber assay and angiogenesis assay were used to examine the metastatic ability, invasiveness and angiogenesis of EC109 cells treated with cyclopamine for 48 h. The expression of Gli-1 mRNA was detected using RT-PCR, and Western blotting was used to examine the protein expressions of Gli-1, matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF).
RESULTSInhibition of the hedgehog signaling pathway by cyclopamine suppressed the migration, invasion, and angiogenesis of EC109 cells. Cyclopamine treatment significantly lowered the expression of Gli-1 mRNA (P<0.05) and the protein expressions of Gli-1, MMP-9 and VEGF (P<0.05).
CONCLUSIONCyclopamine can significantly inhibit the metastatic capacity of EC109 cells possibly by down-regulating MMP-9 and VEGF expression as a result of Gli-1 inhibition.
Cell Line, Tumor ; Esophageal Neoplasms ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; Signal Transduction ; drug effects ; Transcription Factors ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Veratrum Alkaloids ; pharmacology ; Zinc Finger Protein GLI1