Mobile website
Basic Research
FHL2 affects the resistance of glioblastoma U87 cells against temozolomide via MGMT
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To investigate the effect of interfering the expression of four and a half LIM-only protein 2 (FHL2) on the expression of O6-methylguanine DNA methyltransferase (MGMT) and temozolomide (TMZ) resistance of glioblastoma U87 cells.Methods: Lentiviruses carrying different sequences of FHL2 interference sequences (shFHL2-1#, shFHL2-4#) and negative control (shN) were infected into U87 cells, namely shFHL2-1# group, shFHL2-4# group, and shN group, respectively. siMGMT-1#, siMGMT-4#,and siN were transfected into U87 cells by siRNA transfection technology, namely siMGMT-1# group, siMGMT-4# group, and siN group, respectively. The FHL2 or MGMT knockdown efficiency was verified by qPCR and WB. The above groups of cells were treated with TMZ (the DMSO treatment group was used as the control), and then the proliferation of cells with FHL2 or MGMT knockdown before and after TMZ treatment was detected by CCK-8 method and cell clone formation assay, the apoptosis of cells in FHL2 knockdown group before and after TMZ treatment was detected by flow cytometry. WB method and immunofluorescence method was used to detect the effect of FHL2 knockdown on the expression of MGMT in U87 cells. WB method was used to detect the effect of TMZ treatment on the expression levels of FHL2 and MGMT in each group of cells. Results: Glioblastoma U87 cells with FHL2 or MGMT knockdown were successfully constructed. Compared with the shN group, the proliferation ability in cells of shFHL2-1# or shFHL2-4# group was significantly reduced while the apoptosis rate was significantly elevated (all P<0.01), and the expression of MGMT were significantly reduced (all P<0.01). After TMZ treatment, the expression levels of FHL2 and MGMT in the shN group were significantly increased (both P<0.05), while the proliferation and apoptosis of the cells were not significantly changed (all P>0.05) compared with the corresponding DMSO treatment group. The expression levels of FHL2 and MGMT in the cells of shFHL2-1# and shFHL2-4# groups did not change significantly (all P>0.05), but the cell proliferation capacity was significantly reduced, and the apoptosis level was significantly increased (all P<0.01). Knockdown of MGMT slowed down the proliferation of U87 cells in shN group (P<0.01), while the proliferation capacity of cells of siMGMT-1# and siMGMT-4# groups was further reduced after TMZ treatment (all P<0.01). Conclusion: Interfering with FHL2 expression weakened the proliferation ability of U87 cells and increased the apoptosis rate, and downregulated the expression of MGMT, suggesting that FHL2 may regulate the resistance of U87 cells to TMZ by affecting the expression of MGMT.
Keywords:
Project Supported:
Clc Number:
