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【Objective】 To investigate the expression of Kinesin family member 14 (KIF14), and its correlation with clinical prognosis and immune cell infiltration of clear cell renal cell carcinoma (ccRCC). 【Methods】 The correlation between KIF14 expression in ccRCC and different clinicopathological features were analyzed with TCGA, GEO and Ualcan databases. The correlation between KIF14 expression and prognosis was analzyed with Kaplan-Meier method. The correlation between KIF14 expression and immune cell infiltration was analzyed with TIMER. The protein-protein interaction network of KIF14 was conducted with Genemania. The co-expression genes of KIF14 in TCGA-KIRC were picked out in Linkedomics database and were used to perform GO annotations and KEGG pathway enrichment analysis with R software. The biological functions of KIF14 were verified with in vitro functional assay. 【Results】 KIF14 was highly expressed in ccRCC tissue and was positively correlated with clinical stage, pathological grade, and lymphatic metastasis, but negatively correlated with clinical prognosis. KIF14 expression was an independent risk factor for overall survival of ccRCC patients. GO annotations showed that KIF14 was involved in DNA replication, nuclear division, organelle fission, and cell adhesion. KEGG pathway enrichment analysis showed that KIF14 participated in cell cycle and p53 signaling pathway. Genemania analysis indicated KIF14 interacted with CENPE, CIT, KIF23, and other proteins. Timer showed that KIF14 was positively correlated with immune cell infiltration. Knockdown of KIF14 expression suppressed cell proliferation, migration, and invasion of ccRCC. 【Conclusions】 KIF14 may serve as a novel prognostic marker and a potential therapeutic target of clear cell renal cell carcinoma.
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Objective:To investigate the role of kinesin family proteins(KIF)2C in chemoresistance of epithelial ovarian cancer and its potential molecular mechanism.Methods:The differential expression of KIF2C in cisplatin-sensitive cell line A2780 and cisplatin-resistant cell line A2780DDP was detected.Different concentrations and time of cisplatin were added to A2780 cell line to detect the differential expression of KIF2C.The ovarian cancer cell line KIF2C-OV with stable overexpression of KIF2C was constructed by lentivirus infection of A2780DDP cell line.The same concentration of cisplatin was added to KIF2C-OV and its control group(Ctrl-OV)cell lines.The effect of KIF2C overexpression on the proliferation of drug-resistant cell line A2780DDP was detected by Cell Counting Kit-8(CCK-8). Finally, the molecular mechanism of KIF2C resistance in ovarian cancer cell lines was preliminarily explored.Results:KIF2C was significantly down-regulated in A2780DDP cells.After cisplatin was added to A2780 cell line, the expression of KIF2C decreased with the increase of cisplatin concentration and action time.The ovarian cancer cell line KIF2C-OV with stable overexpression of KIF2C was constructed by lentivirus infection of A2780DDP cell line.After adding cisplatin, the proliferation ability of KIF2C-OV cell line was significantly lower than that of Ctrl-OV cell line.The expression of p-Erk was increased in A2780DDP cell line.The expression of p-Erk in A2780 cell line was increased with the increase of cisplatin concentration and time.After overexpression of KIF2C in A2780DDP ovarian cancer cell line, the expression level of p-Erk decreased.Conclusions:KIF2C may be a potential target gene of chemotherapy resistance in ovarian cancer, which may play its role through MAPK/ERK pathway.
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Objective:To analyze the expression and relationship of miR-379 and human kinesin family member 4A (KIF4A) in triple-negative breast cancer (TNBC).Methods:A total of 52 patients diagnosed with TNBC in breast department at Puji Branch of Dongguan People′s Hospital between January 2016 and November 2019 were retrospectively selected as the study group. Meanwhile, 70 patients with non-triple-negative breast cancer (NTNBC) diagnosed in the same period were selected as the control group. The expression levels of miR-379 and KIF4A in both groups were detected. The receiver operating characteristic (ROC) curve was drawn, and the area under the curve (AUC) of miR-379 and KIF4A predicting TNBC was calculated; The relationship between the expression levels of miR-379 and KIF4A and clinicopathological parameters, and the correlation between the expression level of miR-379 and KIF4A were analyzed.Results:The expression level of KIF4A in study group was higher than that in control group [(6.93±0.43) vs (3.75±0.25), P<0.05], and the expression level of miR-379 in study group was lower than that in control group [(0.54±0.17) vs (0.87±0.32), P<0.05]. MiR-379 combined with KIF4A expression had the greatest diagnostic value for TNBC: AUC 0.823 (95% CI: 0.730- 0.917), specificity 0.785, sensitivity 0.950; Univariate analysis showed that there were significant difference in the expression of miR-379 in TNBC patients with different clinical stages, tumor diameter, and lymph node metastasis (all P<0.05); There were significant difference in the expression of KIF4A in TNBC patients with different clinical stages and tumor diameter (all P<0.05). Pearson correlation analysis showed that miR-379 expression was negatively correlated with KIF4A expression in TNBC ( r=-0.349, P<0.05). Conclusions:The expression of miR-379 is down-regulated, while the expression of KIF4A is up-regulated in patients with TNBC. The two are related to poor clinicopathological characteristics, which indicates that they can be used for condition evaluation of the patients.
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Objective:To investigate the expression of silencing kinesin KIF4A in thyroid cancer tissues and its relationship with clinicopathological characteristics of thyroid cancer patients, and to assess the role of KIF4A in the progression of thyroid cancer.Methods:The expression of KIF4A in normal thyroid tissues and the thyroid cancer population and its relationship with disease-free survival of patients were analyzed online by gene expression interaction analysis (GEPIA) database, and the expression of KIF4A in tumor tissues and paraneoplastic tissues of thyroid cancer patients was assessed by immunohistochemical assays. The patients were divided into high- and low-expression groups according to the staining intensity, and the correlation between the expression of KIF4A and clinicopathological features was analyzed. The effect of KIF4A on the proliferation of thyroid cancer cells was explored by a clone formation assay and an MTT assay.Results:According to the analysis of the web-based database, KIF4A showed significantly high expression in human thyroid cancer tissues, and disease-free survival was significantly lower in highly expressed patients. The results of the case analysis showed that the correlation between KIF4A expression intensity and gender, age, and lymph node metastasis in thyroid cancer patients was not statistically significant (all P>0.05), and the correlation with TNM stage and intraglandular dissemination was statistically significant (all P<0.05). The results of the colony formation assay and the MTT assay showed that the expression of KIF4A promoted the proliferation of thyroid cancer cells ( P<0.05). Conclusions:KIF4A can promote the progression of thyroid cancer and has the potential to become a new therapeutic target for thyroid cancer.
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Objective:To analyze the relationship between the expression and prognosis of kinesin family member 4A (KIF4A) in renal clear cell carcinoma and explore its potential mechanism.Methods:Information on the KIF4A gene in renal clear cell carcinoma was retrieved from the UALCAN database, including expression levels, survival analysis, and positive and negative correlation gene data, from which the expression levels, prognostic information, and potential mechanisms of KIF4A in renal clear cell carcinoma were derived.Results:KIF4A is highly expressed in renal clear cell carcinoma, and male patients have a higher expression rate than female patients. The higher the tumor grade and the later the N stage, the more expression ( P<0.05). The survival analysis showed that the expression level and prognosis of KIF4A were negatively correlated ( P<0.05). Cyclin B2 (CCNB2), kinesin family member 23 (KIF23), cell division cycle associated 5 (CDCA5), and KIF4A were significantly positively correlated, whereas DHRS12, crumbs protein homolog 3 (CRB3), β-hydroxybutyrate dehydrogenase 2 (BDH2), and KIF4A were significantly negatively correlated. Conclusions:KIF4A plays an important role in the development of renal clear cell carcinoma and can be used as a potential marker and therapeutic target for the diagnosis and prognosis of renal clear cell carcinoma, including in children.
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Objective:To investigate the expression and clinical significance of kinesin family member 23 (KIF23) in human triple-negative breast cancer (TNBC).Methods:The clinicopathological characteristics of 74 cases of TNBC patients were retrospectively analyzed. The expression of KIF23 in tumor tissues and adjacent normal tissues was detected by immunohistochemical method, and the expression of KIF23 in TNBC patients and its relationship with clinicopathological characteristics were analyzed. The mRNA level of KIF23 in TNBC tissues and normal tissues adjacent to cancer was analyzed by bioinformatics methods, and the relationship between its expression and the survival rate of patients was also analyzed.Results:The results of bioinformatics analysis showed that the high expression of KIF23 in TNBC tissue was significantly related to the overall survival and disease-free survival of patients (all P<0.05). In TNBC tissues, the positive high expression rate of KIF23 was 64.9%, while it was mainly low or no expression in adjacent tissues. The high expression of KIF23 was significantly correlated with the tumor size and pTNM stage of TNBC patients (all P<0.05). Conclusions:KIF23 plays a regulatory role in the progression of TNBC, and it can be used as a new diagnostic and therapeutic target for TNBC.
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Objective:To explore the function and role of kinesin family member C1 (KIFC1) in triple negative breast cancer (TNBC).Methods:The mRNA level of KIFC1 in TNBC tissues and normal tissues adjacent to cancer was analyzed by bioinformatics methods, and the relationship between its expression and the survival rate of thepatients was analyzed. The clinicopathological characteristics of 96 TNBC patients were retrospectively analyzed. Immunohistochemical method was used to detect the expression of KIFC1 in tumor tissues and normal tissues adjacent to cancer, and to analyze the expression of KIFC1 in TNBC patients and its relationship with clinicopathological characteristics.Results:The results of bioinformatics analysis showed that KIFC1 is highly expressed in TNBC tissue and is correlated with the patient's disease-free survival ( P<0.05). In TNBC tissue, the positive high expression rate of KIF23 is 58.3%, while it is mainly low or no expressionin adjacent tissues. The high expression of KIF23 is related to the tumor grade of TNBC patients ( P<0.05). The results of in vitro cell experiments show that knocking down KIFC1 can significantly reduce the colony forming ability and proliferation ability of TNBC cells. The results of in vivo experiments in mice showed that knocking down KIFC1 can significantly reduce tumor volume. Conclusions:KIFC1 can be used as a prognostic factor of TNBC. Low expression of KIFC1 can inhibit the proliferation of TNBC cells in vivo and in vitro. KIFC1 is expected to be a prognostic marker and therapeutic target for TNBC.
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This study aimed to explore the correlation of kinesin family member 2A (KIF2A) expression with disease risk, clinical characteristics, and prognosis of acute myeloid leukemia (AML), and investigate the effect of KIF2A knockdown on AML cell activities in vitro. Bone marrow samples were collected from 176 AML patients and 40 healthy donors, and KIF2A expression was measured by real-time quantitative polymerase chain reaction. Treatment response, event-free survival (EFS), and overall survival (OS) were assessed in AML patients. In vitro, KIF2A expression in AML cell lines and CD34+ cells (from healthy donors) was measured, and the effect of KIF2A knockdown on AML cell proliferation and apoptosis in HL-60 and KG-1 cells was detected. KIF2A expression was greater in AML patients compared to healthy donors, and receiver operating characteristic curve indicated that KIF2A expression predicted increased AML risk (area under curve: 0.793 (95%CI: 0.724-0.826)). In AML patients, KIF2A expression positively correlated with white blood cells, monosomal karyotype, and high risk stratification. Furthermore, no correlation of KIF2A expression with complete remission or hematopoietic stem cell transplantation was found. Kaplan-Meier curves showed that KIF2A expression was negatively correlated with EFS and OS. In vitro experiments showed that KIF2A was overexpressed in AML cell lines (KG-1, HL-60, ME-1, and HT-93) compared to CD34+ cells, moreover, cell proliferation was reduced but apoptosis was increased by KIF2A knockdown in HL-60 and KG-1 cells. In conclusion, KIF2A showed potential to be a biomarker and treatment target in AML.
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Humans , Male , Female , Adult , Middle Aged , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/genetics , Kinesins/genetics , Biomarkers, Tumor/genetics , Survival Rate , Risk Factors , Apoptosis , HL-60 Cells , Cell Proliferation , Gene Knockdown TechniquesABSTRACT
Objective:To investigate the induction of specific T lymphocyte by bispecific monoclonal antibody in pancreatic cancer and its killing effects on KIF20A positive pancreatic cancer PANC1 cell line.Methods:CD 3/KIF20A bispecific monoclonal antibody was prepared and concentrated by chemical cross-linking method and purified by Sephrose-25 gel chromatography. Peripheral blood samples of healthy volunteers were collected, and monocytes were isolated using lymphocyte separation solution, and then cultured as dendritic cells (DC) and T cells respectively, and then co-cultured as DC-T cells. Meanwhile vitamin C was used to treat DC-T cells (vcDC-T cells). The levels of IFN-γ, IL-2, IL-4 and IL-12 in the supernatants and T cell subsets were detected by flow cytometry. About 1×10 5 T cells, DC-T cells, and vcDC-T cells with 10, 50, 100 and 300 ng CD 3/KIF20A antibody loaded were cocultured with PANC1 cells (20∶1) for 2, 6 and 10 hours to determine the highest killing rate dosage of CD 3/KIF20A antibody loaded cells. DC-T cells and DC-T cells, vcDC-T cells loaded with the highest killing rate dosage of CD 3/KIF20A antibody were cocultured with PANC1 cells (20∶1) for 2, 6, 8, 10 and 12 hours. The aggregation effect of effector cells on target cells was observed under inverted microscope, the killing rate of tumor cells was detected by LDH method. Results:The molecular weight of CD 3/KIF20A antibody was 130 000 measured and validated by SDS gel electrophoresis. The ratio of CD 8+ CD 28+ and CD40L subsets of vcDC-T cells was increased [(47.6±15.8)% vs (38.2±7.6)%, (52.1±4.9)% vs (44.7±3.2)% ] compared with that of DC-T cells, the ratio of negative regulatory cells (Treg) was decreased [(4.3±0.8)% vs (8.3±1.1)%]; the release of IL-2, IFN-γ and IL-12 was increased [(201.2±17.3) ng/L, (163.4±13.1)ng/L, (303.3±22.6)ng/L vs 221.8±17.6)ng/L, (190.4±11.7)ng/L vs (80.3±8.6)ng/L]. All the differences were statistically significant ( P<0.01). 100 ng CD 3/KIF20A loaded T cells were observed under microscope, which obviously targeted KIF20A + pancreatic cancer PANC1 cells and had a strongest killing power. At the killing cells to targeting cells ratio of 20∶1 with 4-hour coculture, the killing rate of CD 3/KIF20A-vcDC-T cells on PANC1 cells was (88.6±2.6)%, which was significantly higher than (68.4±3.4)% and (39.2±2.1)% in the CD3/KIF20A-DC-T group and (39.2±2.1)% in the DC-T group, increasing by 20% and at lease 45%, respectively. Conclusions:DC-T cells loaded with CD 3/KIF20A antibody can significantly increase the killing rate of KIF20A positive pancreatic cancer PANC1 cells, and vitamin C intervention can further enhance the killing ability of antibody loaded T cells.
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Animals , Humans , Mice , beta Catenin , Blotting, Western , Breast Neoplasms , Breast , Cell Self Renewal , Flow Cytometry , Fluorescent Antibody Technique , Genome , In Vitro Techniques , Kinesins , Luciferases , Neoplasm Metastasis , Neoplastic Stem Cells , Prognosis , Real-Time Polymerase Chain Reaction , Stem CellsABSTRACT
Objective To study the expression of kinesin family member C1 ( KIFC1 ) in hepatocellular carcinoma (HCC) and analyze its correlation with the clinicopathological features and prognosis of patients. Methods The expression levels of KIFC1 protein in the HCC tissues from 82 patients were determined by immunohistochemical staining. The correlation between KIFC1 protein and clinicopathological characteristics (including age, gender, tumor nodules, tumor grade, tumor volume, lymph node metastasis, and alpha-fetoprotein expression) was analyzed. The Kaplan-Meier analysis was used to analyze the effect of KIFC1 expression level on overall survival and progression-free survival in patients with HCC. The expression level of KIFC1 mRNA in liver cancer tissue was analyzed by GPEIA database. The correlation between KIFC1 expression and prognosis was analyzed by KM-plotter. Results KIFC1 protein is significantly overexpressed in liver cancer tissues, and its expression level is significantly correlated with tumor nodule number (P=0.023) and tumor size (P=0.011). Patients with high expression of KIFC1 had poor overall disease and disease-free survival (all P<0.05). KIFC1 mRNA is significantly overexpressed in liver cancer tissues and correlated with disease-free survival and overall survival. Conclusions The expression of KIFC1 protein is highly expressed in liver cancer tissues, and its expression level is related to the clinicopathological characteristics of liver cancer. Bioinformatics analysis results show that KIFC1 is related to the poor prognosis of patients, suggesting that KIFC1 is expected to be a potential predictor and therapeutic target for liver cancer prognosis.
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Objective To explore the expression of kinesin family member 20A (KIF20A) in hepatocellular carcinoma (HCC) and its prognostic significance.Methods By using bioinformatics methods in Gene Expression Profiling Interactive Analysis (GEPIA),Oncomine and The Human Protein Atlas (THPA) online analysis websites,the mRNA and protein expression information of KIF20A in HCC was analyzed based on the large cancer public data including The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO).The Kaplan-Meier method was used to perform patients' survival analysis based on TCGA liver cancer data,and the survival rates were compared by log-rank method.Pearson correlation analysis was performed to investigate the expression of KIF20A and some key molecules in phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathways.Results GEPIA retrieved 369 cases of HCC and 50 cases of normal liver tissue containing KIF20A mRNA expression information.Oncomine retrieved a total of 4 studies on KIF20A mRNA in HCC.All results showed that compared with the normal liver tissues,the mRNA expression level of KIF20A was significantly higher in HCC tissues (P ＜ 0.001;t =8.766,P ＜ 0.001;t =24.329,P ＜ 0.001;t =7.398,P ＜0.001;t =3.191,P =0.001).Besides,THPA online analysis websites showed that KIF20A protein was low or not expressed in normal liver tissues,but it was significantly higher in HCC tissues,and this result was consistent with the mRNA analysis result.Moreover,the survival analysis found that the expression of KIF20A was correlated with the overall survival (OS) and disease-free survival (DFS) of HCC patients,and the prognosis of patients with high KIF20A expression was poor (P =0.003;P ＜ 0.001).Additionally,further correlation analysis found that KIF20A gene was positively correlated with phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA),AKT1,mammalian target of rapamycin (mTOR),hypoxia-inducible factor 1ot (HIF1A) and vascular endothelial growth factor A (VEGFA) genes in HCC (R =0.43,P ＜ 0.001;R =0.29,P＜0.001;R=0.18,P＜0.001;R=0.39,P＜0.001;R=0.37,P＜0.001).Conclusion Bioinformatics analysis results indicate that KIF20A is highly expressed in HCC tissues and KIF20A is associated with the prognosis of HCC patients,the mechanism of which may be related to the regulation of PI3K/AKT signaling pathway.It is worth further study in the future.
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Objective To evaluate the role of spinal kinesin superfamily motor protein 17 (KIF17)in remifentanil-induced hyperalgesia in rats with incisional pain. Methods Forty male Sprague-Dawley rats, aged 2-3 months, weighing 240-260 g, were divided into 5 groups(n=8 each)using a random number table: control group(group C), remifentanil group(group R), incisional pain group (group I), remifentanil plus incisional pain group(group R+I)and remifentanil plus incisional pain plus KIF17 inhibitor group(group R+I+M). Normal saline 15 ml was intravenously infused for 60 min in group C. Remifentanil 1 μg·kg-1·min-1was intravenously infused for 60 min in group R. In group I, in-cisional pain model was established, and normal saline 15 ml was intravenously infused for 60 min at the same time. In group R+I, incisional pain model was established, and remifentanil 15 ml was intravenous-ly infused for 60 min at the same time. In group R+I+M, KIF17 inhibitor Myr-Rc-13 10 μg(in 10 μl dimethyl sulfoxide)was injected intrathecally, and remifentanil 1 μg·kg-1·min-1was intravenously in-fused for 60 min while incisional pain model was established. The mechanical paw withdrawal threshold (MWT)and thermal paw withdrawal latency(TWL)were measured at 24 h before remifentanil or normal saline infusion and at 2, 6, 24 and 48 h after the end of infusion. The rats were sacrificed after the last measurement of pain threshold, and the lumbar segment of the spinal cord was removed for determination of the expression of KIF17 and phosphorylated KIF17(pKIF17)by Western blot. Results Compared with group C, MWT was significantly decreased, TWL was shortened, and the expression of KIF17 and pKIF17 was up-regulated in R, I and R+I groups(P<005). Compared with R and I groups, MWT was signifi-cantly decreased, TWL was shortened, and the expression of KIF17 and pKIF17 was up-regulated in group R+I(P<005). Compared with group I+R, MWT was significantly increased, TWL was prolonged, and the expression of KIF17 and pKIF17 was down-regulated in group R+I+M(P<005). Conclusion In-creased activity of KIF17 is involved in the development and maintenance of remifentanil-induced hyperalge-sia in rats with incisional pain.
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Pterocarpanos representam a maior classe de isoflavonóides, depois das isoflavonase estudos recentes tem revelado que os representantes deste grupo podem agir em alvos específicos da mitose. O presente estudo avaliou o potencial citotóxico, genotóxico e mutagênico, do composto CP001,em célulasnormais e linhagens tumorais humanas, além do efeito inibitório sobre a quinesina Eg5fazendo uso de ensaios in vitro e com cálculos de bioquímica quântica.Os ensaios de citotoxicidade mostraram que o CP001apresentou efeito significativo sobre as linhagens tumorais testadas (HL-60, HCT-116, OVCAR-8, SF-295)com IC50 variandoentre 0,2 e 3,61 μM. Efeito confirmado na curva de crescimento cinético em tempo real, onde o CP001inibiu o crescimento da linhagem OVCAR-8 na concentração de 4 μM, semelhante ao paclitaxel (0,5 μM).Desta forma, a fim de determinar o mecanismo de ação envolvido, uma sequência de experimentos in vitroforam realizados.A avaliação do conteúdo de DNA nuclear foi mensurada em células OVCAR-8, para analisar o efeito do pterocarpano sobre as fases do ciclo celular, revelando que o CP001 é capaz de parar o ciclo celular na fase G2/M, na concetração de 5 μM, e com efeito potencializador, quando colocado em conjunto com o paclitaxel. A parada do ciclo celular na fase G2/M pode estar relacionado a ação do composto na tubulina. O ensaio da polimerização da tubulina foi conduzido e mostrou-se que o CP001 possui velocidade de polimerização (Vma = 80.95 mOD/min), inferior a do paclitaxel (Vmax = 100 mDO/min) e próxima a do monastrol (Vmax = 88.46 mOD/min). Estes dados mostram que a interferência na polimerização da tubulina não é tão significativa quanto aquela apresentada no paclitaxel. A ação em proteínas específicas da mitose foi outra possibilidade testada...
Pterocarpans represented the largest isoflavonoid class after isoflavones and recent studies have revealed that representatives of this group can act on specific targets of mitosis.This study evaluated the potential cytotoxic , genotoxic and mutagenic of the CP001 in tumoral and normal human cell lines. The inhibitory effect in Eg5 kinesin and quantum biochemistry calculation has been peformed as well. Cytotoxicity tests showed that CP001 hads ignificant effect on the tested tumoral lines (HL-60, HCT-116, OVCAR-8, SF-295) with IC50 ranging between 0.2 and 3.61 μ M. Effect confirmed bykinetic growth curve in real time, where the CP001inhibit growth of OVCAR-8 cell line at a concentration of 4 uM , similar to paclitaxel (0.5 mM). Thus, in order to determine the mechanism of action involved a sequence of in vitro experiments were per formed .The assessment of nuclear DNA content was measured in OVCAR-8 cells to examine the effect of pterocarpanon the cellcycle phases , showing that CP001is capable to arrest cell cycle at the G2/ M phase, concentration of 5 uM , and potentiating effect when added in combination with paclitaxel.The cell cycle arrest in G2/M phase may be related to action of the compound on tubulin.The tubulin assay polymerization was conducted and revealed that CP001 has polymerization rate (Vmax = 80.95 mOD / min ) b elow paclitaxel(Vmax = 100 mOD / min) and close tomonastrol ( Vmax = 88.46 mOD / min).These data suggest that interference with tubulin polymerization is not as significant as that shown in paclitaxel.Specific action mitosis proteins was another possibility tested...
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Humans , Neoplasms , Molecular Docking Simulation , KinesinsABSTRACT
Copy number variations at multiple chromosomal loci, including 16p11.2, have recently been implicated in the pathogenesis of autism spectrum disorder (ASD), a neurodevelopmental disease that affects 1~3% of children worldwide. The aim of this study was to investigate the roles of human genes at the 16p11.2 loci in synaptic development using Drosophila larval neuromuscular junctions (NMJ), a well-established model synapse with stereotypic innervation patterns. We conducted a preliminary genetic screen based on RNA interference in combination with the GAL4-UAS system, followed by mutational analyses. Our result indicated that disruption of klp68D, a gene closely related to human KIF22, caused ectopic innervations of axon branches forming type III boutons in muscle 13, along with less frequent re-routing of other axon branches. In addition, mutations in klp64D, of which gene product forms Kinesin-2 complex with KLP68D, led to similar targeting errors of type III axons. Mutant phenotypes were at least partially reproduced by knockdown of each gene via RNA interference. Taken together, our data suggest the roles of Kinesin-2 proteins, including KLP68D and KLP64D, in ensuring proper synaptic wiring.
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Child , Humans , Autistic Disorder , Axons , Drosophila , Genes, vif , Neuromuscular Junction , Phenotype , RNA Interference , Synapses , Autism Spectrum DisorderABSTRACT
Kinesin-2 family proteins, including KIF3A, KIF3B, KIF3C and KIF17, are members of the kinesin superfamily motor proteins , which transport various proteins and vesicles in the cell and play diverse biological functions . Recently, studies on members of kinesin-2 family proteins suggest that they play fundamental roles during ciliary transport , whose defects can lead to abnormal cilia development , the major cause of human ciliopathies .In this review , we will sum-marize the functions of this motor protein family during ciliogenesis and focus mainly on their roles in the development of model organisms .
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Kinesin spindle protein ( KSP) inhibitors is an important direction for the discovery of anticancer agents. Several KSP in-hibitors have been studied in clinic trials. The discovery of ATP-competitive KSP inhibitors may be a new approach for searching novel a-gents to overcome the mutation-mediated resistance to the allosteric inhibitors. The progress in the discovery of ATP-competitive KSP in-hibitors was reviewed in the paper to provide reference for the further development of KSP inhibitors.
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Objective To explore the role of spinal monocyte chemoattractant protein?1 ( MCP?1) ?extracellular signal?regulated protein kinase ( ERK)?kinesin superfamily motor protein 17 ( KIF17)∕N?methyl?D?aspartate receptor subunit 2B ( NR2B) signaling pathway in the maintenance of type 2 diabetic neuropathic pain (DNP) in rats. Methods Type 2 diabetes mellitus was induced by a high?fat and high?sucrose diet and intraperitoneal streptozotocin ( STZ) 35 mg∕kg, and confirmed by fasting blood glucose level≥16?7 mmol∕L 3 days later in male Sprague?Dawley rats aged 6 weeks. Type 2 DNP was confirmed when the mechanical paw withdrawal threshold ( MWT ) and thermal paw withdrawl latency ( TWL ) measured on day 14 after STZ administration decreased to< 80% of the baseline value. The rats with type 2 DNP were randomly divided into 4 groups ( n=36 each) using a random number table: type 2 DNP group (group DNP), type 2 DNP +MCP?1 neutralizing antibody group (group DM), type 2 DNP +ERK inhibi?tor group (group DE) and type 2 DNP + dimethyl sulfoxide group ( group DD). In DM, DE and DD groups, 0?1 ng∕μl MCP?1 neutralizing antibody 10 μl, 0?5 μg∕μl U0126 10 μl and 5 % dimethyl sulfoxide 10 μl were injected intrathecally, respectively, once a day for 14 consecutive days starting from 14 days after administration of STZ. Another 36 normal rats fed a common forage diet were adopted as con?trol group ( group C) . MWT and TWL were measured before STZ injection and at 1, 3, 7 and 14 days after STZ injection ( T0-4 ) . Nine rats were sacrificed after measurement of pain thresholds at T1-4 , and the lumbar segments ( L4-6 ) of the spinal cord were removed for determination of the expression of phosphoryla?ted ERK (p?ERK), KIF17 and phosphorylated NR2B (p?NR2B) by Western blot. Results Compared with group C, the MWT was significantly decreased, the TWL was shortened, and the expression of p?ERK, KIF17 and p?NR2B was up?regulated at T1-4 in DNP, DM, DE and DD groups. Compared with group DNP, the MWT at T3-4 in group DM and at T2-4 in group DE was significantly increased, the TWL at T3-4 in group DM and at T2-4 in group DE was prolonged, and the expression of p?ERK, KIF17 and p?NR2B was down?regulated at T2-4 in DM and DE groups, and no significant changes were found in the pa?rameters mentioned above in group DD. Conclusion Spinal MCP?1?ERK?KIF17∕NR2B signaling pathway is involved in the maintenance of type 2 DNP in rats.
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Objective To investigate the effects of repeated intrathecally kinesin superfamily protein 17 (KIF17) antisense oligodeoxynucleotide (ODN) on the expression of mLin10 and NR2B in spinal cord in a mouse model of bone cancer pain.Methods Fifty-six male C3H/HeJ mice,aged 4 ～ 6 weeks,weighting 20 ～ 25 g,were randomly divided into two groups:sham operation group (group S,n=20) and bone cancer pain group (group T,n=36).20μl α-minimal essence medium (α-MEM) which containing 2× 105 NCTC2472 osteosarcoma cells was injected into the intramedullary space of the right femur in group T.In group S,no cancer cell was instead.The number of spontaneous flinches (NSF) and the paw withdrawal mechanical threshold (PWMT) were measured at the day before (base) and the days 4,7,10 and 14 after inoculation.According to the corresponding time points,twenty-four mice were sacrificed for determination the expression of KIF17,mLin10 and NR2B using Western blot.Then,the mice of group T were randomly divided into three groups (n=8,T1,T2,T3,group).In group S and group T1,Saline 5 μl was injected intrathecally.KIF17 sense ODN and antisense ODN,5 μg/5μl were respectively injected in group T2 and T3 for 6 consecutive days.Pain behaviors were assessed at the days 2-6 after the first injection.And determinated the KIF17,mLin10 and NR2B expression,again.Results Compared with group S,the NSF was increased and the PWMT was decreased at the days 7,10 and 14 after inoculation in group T (P＜0.05).Compared with the base ((0.65±0.15),(1.06±0.06),(1.01±0.14)),the expression of KIF17,mLin10 and NR2B (14d:(1.13 ±0.06),(2.17 ± 0.37),(1.85 ± 0.32)) were increased at the days 7,10 and 14 after inoculation in group T(P＜0.05).During the course of the injection,compared with group T1 and T2,the NSF was decreased and the PWMT was increased significantly in the group T3(P＜0.05),the expression of KIF17,mLin10 and NR2B((0.88±0.08),(0.96±0.11),(1.03±0.08)) were reduced in group T3 (P＜0.05).Conclusion Intrathecal KIF 17 antisense ODN in the mice of bone cancer pain improves the pain behaviors,and inhibits the up-regulated of KIF17,mLin10 and NR2B during the course of the injection.
ABSTRACT
BACKGROUND: Vascular endothelial growth factor (VEGF) is involved in the growth of new blood vessels that feed tumors and kinesin spindle protein (KSP) plays a critical role in mitosis involving in cell proliferation. Simultaneous silencing of VEGF and KSP, an attractive and viable approach in cancer, leads on restricting cancer progression. The purpose of this study is to examine the therapeutic potential of dual gene targeted siRNA cocktail on human hepatocellular carcinoma Hep3B cells. RESULTS: The predesigned siRNAs could inhibit VEGF and KSP at mRNA level. siRNA cocktail showed a further downregulation on KSP mRNA and protein levels compared to KSP-siRNA or VEGF-siRNA, but not on VEGF expression. It also exhibited greater suppression on cell proliferation as well as cell migration or invasion capabilities and induction of apoptosis in Hep3B cells than single siRNA simultaneously. This could be explained by the significant downregulation of Cyclin D1, Bcl-2 and Survivin. However, no sigificant difference in the mRNA and protein levels of ANG2, involving inhibition of angiogenesis was found in HUVECs cultured with supernatant of Hep3B cells treated with siRNA cocktail, compared to that of VEGF-siRNA. CONCLUSION: Silencing of VEGF and KSP plays a key role in inhibiting cell proliferation, migration, invasion and inducing apoptosis of Hep3B cells. Simultaneous silencing of VEGF and KSP using siRNA cocktail yields promising results for eradicating hepatocellular carcinoma cells, a new direction for liver cancer treatment.