Journal: BMC Urology

Article Title: Elevated MTA1 induced the migration and invasion of renal cell carcinoma through the NF-κB pathway

doi: 10.1186/s12894-020-00731-1

Figure Lengend Snippet: MTA1 regulated the expression of metastasis-related factors via the NFκB pathway. A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. a Protein levels of MTA1; cells were collected for WB with MTA1 and GAPDH antibodies. The bands of over-expressed MTA1 were from Additional file : Fig. 2A and down-expressed MTA1 were from Additional file : Fig. 2B with arrow. b Cells were lysed and used in qRT-PCR assays to measure the E-cadherin mRNA expression. Statistical analysis was performed using Student’s t test. b , c A498 cells were transfected with pcDNA3.1-Flag and Flag-MTA1. After 36 h, the Flag-MTA1 group was treated with 10 nM PDTC for 12 h. Then cells were harvested for qRT-PCR evaluation to measure the mRNA expression of MMP2 ( b ) and MMP9 ( c ). Statistical analysis was performed using ANOVA. d A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. After 48 h, cells were subjected to western blotting analysis using antibodies targeting p-p65, p65 and ACTB. The bands of p-p65, p65 and ACTB were from Additional file : Fig. 3. e, f Image J was used to calculate the gray scanned bands of p-p65 ( b ) and p65 (C). * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001; NS not significant

Article Snippet: The human kidney cancer cell line A498 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot

Journal: BMC Urology

Article Title: Elevated MTA1 induced the migration and invasion of renal cell carcinoma through the NF-κB pathway

doi: 10.1186/s12894-020-00731-1

Figure Lengend Snippet: MTA1 promotes A498 cell migration. A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. After 24 h, cells in the four conditions were subjected to the wound healing assay. Macrographs were taken under ×100 magnification

Article Snippet: The human kidney cancer cell line A498 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, Transfection, Wound Healing Assay

Journal: BMC Urology

Article Title: Elevated MTA1 induced the migration and invasion of renal cell carcinoma through the NF-κB pathway

doi: 10.1186/s12894-020-00731-1

Figure Lengend Snippet: MTA1 enhances the invasion of A498 cells. a A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, siNC, and si-MTA1. After 24 h, the transwell cell invasion assay using A498 cells was performed, and macrographs were taken under ×40 magnification. Scale bar: 50 µm. b Image J was used to count the transmigrated cells and Student’s t test to analyze differences. ** p < 0.01; *** p < 0.001

Article Snippet: The human kidney cancer cell line A498 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Transfection, Invasion Assay

Journal: BMC Urology

Article Title: Elevated MTA1 induced the migration and invasion of renal cell carcinoma through the NF-κB pathway

doi: 10.1186/s12894-020-00731-1

Figure Lengend Snippet: MTA1 regulated the migration and invasion of RCC cells via NF-κB. A498 cells were transfected with pcDNA3.1-Flag, Flag-MTA1, and Flag-MTA1 + PDTC (10 nM). a In the wound healing assay, MTA1 improved the migration of RCC cells, while the addition of the inhibitor PDTC blocked the effect of MTA1 on migration. Scale bar: 100×. b In the transwell assay, MTA1 markedly induced invasion of A498 cells, but compared to the MTA1 group, the invasion of MTA1 + PDTC A498 cells was lower. ** p < 0.01; *** p < 0.001; Scale bar: ×40

Article Snippet: The human kidney cancer cell line A498 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, Transfection, Wound Healing Assay, Transwell Assay

Journal: BMC Urology

Article Title: Elevated MTA1 induced the migration and invasion of renal cell carcinoma through the NF-κB pathway

doi: 10.1186/s12894-020-00731-1

Figure Lengend Snippet: The expression of MTA1 is up-regulated in RCCs. a Immunohistochemistry was used to characterize the expression of MTA1 in RCC and adjacent tissues. MTA1 was highly expressed in ccRCCs, compared to very weak staining in adjacent tissue. b Image Pro Plus was used for the statistical analysis of the positive signal (IOD) of MTA1 expression in ccRCCs and the adjacent tissue. The expression of MTA1 was significantly up-regulated in ccRCCs. ** p < 0.01; scale bar: 100 µm and 50 µm. c The expression of MTA1 in RCC cell lines. Normal renal cell line HEK293T and RCC cell lines A498 and 768-O cell lysates were used in western blotting analysis with MTA1 and GAPDH antibodies. The bands of MTA1 were from Additional file : Fig. 1 and gapdh were from Additional file : Fig. 1

Article Snippet: The human kidney cancer cell line A498 was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: The high expression of circ-ACTR2 was detected in DN samples and HG-treated HRMCs. A , B The qRT-PCR was performed to detect the circ-ACTR2 level in normal and DN samples ( A ) or control and HG-treated HRMCs ( B ). C The determination of circ-ACTR2 and linear ACTR2 was conducted by qRT-PCR after treatment of RNase R in total RNA. D The circ-ACTR2, U6 and GAPDH levels were assayed by qRT-PCR in cytoplasm and nucleus. ** P < 0.01, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: HG-induced cell damages in HRMCs were partly reversed by the silence of circ-ACTR2. A The expression of circ-ACTR2 was quantified using qRT-PCR in four groups of control, HG, HG + si-NC, HG + si-ACTR2 in HRMCs. B Cell viability detection was performed by CCK-8 assay. C Inflammatory cytokines IL-6 and TNF-α were determined by ELISA. D Cell proliferation analysis was performed by EdU assay. E The protein examination of collagen I and collagen IV was conducted by western blot. F , G SOD activity ( F ) and MDA level ( G ) were respectively measured using the corresponding kits. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Quantitative RT-PCR, Control, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, EdU Assay, Western Blot, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2 exhibited the sponge function of miR-205-5p. A Starbase v2.0 predicted the binding sites between circ-ACTR2 and miR-205-5p. B The miR-205-5p level was examined by qRT-PCR after transfection of miR-NC or miR-205-5p. C , D Dual-luciferase reporter assay ( C ) and RIP assay ( D ) were used to confirm whether circ-ACTR2 combined with miR-205-5p. E The quantification of miR-205-5p was carried out using qRT-PCR in normal and DN tissues. F The linear analysis between circ-ACTR2 and miR-205-5p was performed using the Pearson’s correlation coefficient. G The effect of HG on the miR-205-5p expression was analyzed via qRT-PCR. H The level of circ-ACTR2 was detected by qRT-PCR in control, HG, HG + pcD5-ciR or HG + circ-ACTR2 group. I The regulation of circ-ACTR2 inhibition or overexpression on the miR-205-5p expression was performed by qRT-PCR in HG-treated HRMCs. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Binding Assay, Quantitative RT-PCR, Transfection, Luciferase, Reporter Assay, Expressing, Control, Inhibition, Over Expression

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2/miR-205-5p axis affected the regulation of HG in HRMCs. A The transfection efficiency of anti-miR-205-5p was assessed by qRT-PCR. B The qRT-PCR was applied for the expression detection of miR-205-5p after transfection of si-NC, si-ACTR2, si-ACTR2 + anti-miR-NC or si-ACTR2 + anti-miR-205-5p in HG-treated HRMCs. C Cell viability was examined via CCK-8 assay. D The concentrations of IL-6 and TNF-α were tested via ELSIA. E Cell proliferation was assessed via EdU assay. F , G The levels of collagen I and collagen IV were assayed via western blot. H , I Oxidative stress was evaluated by SOD activity ( H ) and MDA level ( I ) using the kits. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, EdU Assay, Western Blot, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: HMGA2 downregulation protected against the HG-induced cell dysfunction in HRMCs. A The protein expression of HMGA2 in control, HG, HG + si-NC or HG + si-HMGA2 group was evaluated by western blot. B CCK-8 was performed to analyze cell viability. C ELISA was performed to measure the concentrations of IL-6 and TNF-α. D EdU assay was conducted to determine cell proliferation. E , F Western blot was conducted to detect the protein levels of collagen I and collagen IV. G , H SOD activity ( G ) and MDA level ( H ) by the detection kits were exploited to examine the oxidative stress. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Control, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, EdU Assay, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2 regulated the HMGA2 level by acting as a sponge of miR-205-5p. A , B The mRNA ( A ) and protein ( B ) levels of HMGA2 were respectively detected by qRT-PCR and western blot after transfection of si-NC, si-circ-ACTR2, si-circ-ACTR2 + anti-miR-NC or si-circ-ACTR2 + anti-miR-205-5p in HG-treated HRMCs. ** P < 0.01, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Quantitative RT-PCR, Western Blot, Transfection