ABSTRACT
【Objective】 To explore the role of environmental enrichment (EE) in paternal stress-induced anxiety and depression-like behaviors in offspring and its potential mechanisms. 【Methods】 Male C57BL/6 mice (F0) were treated with unpredictable chronic mild stress (UCMS) and subsequently mated with normal females to obtain F1 offspring mice. The standard environment (SE) and enriched environment (EE) were administered to F1-UCMS offspring mice during their early life (3-5 weeks of age). Anxiety-like behaviors were detected by open field test (OFT) and elevated plus-maze test (EPM); depression-like behaviors were detected via forced swimming test (FST) and sucrose preference test (SPT) at the age of 8 weeks. The expressions of LIM and SH3 domain protein 1 (LASP1) in the hippocampus of adult F1 offspring mice were detected by Real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting. 【Results】 Compared to F1 offspring of normal paternal (F1-Nor), F1 offspring mice of the stressed paternal (F1-UCMS) showed significantly anxiety-like behavior with reduced percentage of time spent in the central region of OFT and in the open arm of EPM (P<0.05); mice from the F1-UCMS group showed a significantly increased percentage of immobility in FST and a reduced percentage of sugar consumption in SPT (P<0.01), which demonstrated significant depression-like behaviors. Compared to the SE group, mice in the EE group had an increased percentage of time spent in the central region of the OFT [males: (7.44±0.75)% vs. (14.93±1.74)%, P<0.01; females: (8.89±1.06)% vs. (15.10±1.82)%, P<0.05] and an increased percentage of time in the open arm of EPM [males: (8.09±1.05)% vs. (14.15±1.88)%, P<0.05; females: (9.13±1.14)% vs. (14.04±1.37)%, P<0.05]. This indicated that EE ameliorated anxiety-like behavior in F1-UCMS mice with paternal stress. Compared to the SE group, mice in the EE group had an decreased percentage of immobility in FST [males: (58.63±4.51)% vs. (42.15±3.81)%, P<0.05; females: (57.96±4.19)% vs. (43.25±4.22)%, P<0.05] and an increased percentage of sugar consumption in SPT [males: (50.38±3.47)% vs. (70.39±3.12)%, P<0.01; females: (52.42±2.84)% vs. (69.99±3.55)%, P<0.01]. This indicated that EE ameliorated depression-like behavior in F1-UCMS mice with paternal stress. Hippocampal LASP1 expression was reduced in SE group compared to F1-Nor group (males: P<0.01; females: P<0.05), while LASP1 was increased in EE group compared to SE group (P<0.05) detected by RT-qPCR and Western blotting. 【Conclusion】 EE ameliorates paternal stress-induced anxiety and depression-like behaviors in F1-UCMS mice, and the mechanism may be associated with increased hippocampal LASP1 expression in F1-UCMS mice.
ABSTRACT
Objective To explore the effects and mechanism of LASP1 gene expression on the proliferation, migration, and invasion of human colorectal cancer (LOVO) cells. Methods LASP1 overexpression plasmids and LASP1 interference plasmids were constructed and transfected to LOVO cells. qRT-PCR was used to detect LASP1 mRNA expression and validate the transfection. MTT method and Tunel staining were used to detect cell proliferation and apoptosis, respectively, and scratch test and Transwell test were employed to determine the migration and invasion abilities of cells. Western blot was applied to analyze the expression of LASP1, p-FAK/FAK, and p-AKT/AKT protein in cells. Results The plasmids were successfully transfected. LASP1 overexpression increased the proliferation, migration, and invasion of LOVO cells, decreased the apoptosis, and increased LASP1, p-FAK/FAK, p-AKT/AKT protein expression (P < 0.01). LASP1 knockdown reduced the proliferation, migration, and invasion of LOVO cells, increased the apoptosis, and decreased LASP1, p-FAK/FAK, and p-AKT/AKT protein expression (P < 0.01). Conclusion LASP1 positively regulates the FAK/AKT signaling pathway to promote the proliferation, migration, and invasion of LOVO cells.
ABSTRACT
Objective To investigate the impacts of LASP-1 on the behavior and cell proliferation ability of tumor derived osteoblast-like cells and non tumor derived osteoblasts. Methods QRT-PCR and Western blot were performed to detect LASP-1 expressions in mouse bone marrow mesenchymal stem cells(BMMSC),human osteoblasts (hOB) and human osteosarcoma cell (MG-63). With the construction of recombinant vector and the transfection of shLASP-1,the expression changes in both LASP-1 and osteocalcin(OCN),and the cell number of cell-cycle phase distribution were analyzed. Results With up-or down-regulation of LASP-1,osteocalcin(OCN) expression,cell proliferation ability and the cell number of G2/M phase in non tumor cells(BMMSC,hOB)had no obvious changes. However,as for the tumor cells(MG-63),with an increased expression of LASP-1,the expression of OCN,cell proliferation number and the cell number at G2/M phase changed. There was no significant effect on the moving ability of each cell. Conclusions LASP-1 has no significant effect on the osteogenic proper-ties,cell proliferation ability and cell cycle distribution of non tumor derived cells,but has a regulatory effect on tumor derived cells(MG-63).
ABSTRACT
Objective@#To investigate the expression of LIM and SH3 protein 1 (LASP1) in renal cell carcinoma and its significance in the invasion and migration of renal clear cell carcinoma 786-O cell line.@*Methods@#The expression level of LASP1 in 41 cases of renal cell carcinoma tissues and normal renal tissues was analyzed by immunohistochemistry. The relationship between the expression level of LASP1 and clinical characteristics was further analyzed. Expression of LASP1 in 10 cases of tumor tissues with or without lymph node metastasis was analyzed by Western blot. Furthermore, small interfering RNA (siRNA) targeting LASP1 was constructed and transfected into 786-O cells to downregulate LASP1 expression. The interference effect of LASP1 siRNA on LASP1 protein and the expression of related proteins in epithelial mesenchymal transition (EMT) pathway were detected by Western blot. The effects of LASP1 knockdown on cell proliferation, migration and invasion and gene expression were then assessed using CCK8 assay, transwell cell migration system and western blot analysis, respectively.@*Results@#The positive rate of LASP1 expression in renal clear cell carcinoma tissues was 90.2% (37/41), which was significantly higher than that in the adjacent tissues (29.3%, P=0.002). The expression of LASP1 in renal cell carcinoma was positively correlated with lymph node metastasis and TNM stage of renal cell carcinoma (P<0.05). The results of Western blot showed that LASP1 (0.696±0.053) was highly expressed in renal cell carcinoma (1.459±0.628), especially in cases with lymph node metastasis (2.692±0.186, P<0.05). The LASP1 siRNA remarkably down-regulated the expression of LASP1 protein in 786-O cells. The abilities of proliferation, invasion and migration of 786-O cells were decreased significantly in the LASP1 siRNA groups.The relative expression of E-cadherin protein in the siRNA group (0.848±0.020) was significantly higher than those in the siRNA-NC group (0.671±0.018) and control group (0.691±0.037, P<0.05). The relative expression of N-cadherin protein in the siRNA group (0.449±0.047) was significantly lower than those in the siRNA-NC group (0.613±0.018) and control group (0.633±0.045, P<0.05). The relative expression of vimentin protein in the siRNA group (0.477±0.029) was significantly lower than those in the siRNA-NC group (0.598±0.069) and control group (0.633±0.045, P<0.05 for both).@*Conclusions@#LASP1 is highly expressed in renal clear cell carcinoma, which is closely related to the development of the cancer. The effects of LASP1 on the invasion and migration of 786-O cells and lymph node metastasis may be related to the EMT.