FBXW7α对结直肠癌细胞中热休克转录因子1表达和定位的影响

孙傲; 焦寒; 付杰军

doi:10.3971/j.issn.1000-8578.2021.20.1216

FBXW7α对结直肠癌细胞中热休克转录因子1表达和定位的影响

Effects of FBXW7α on Expression and Localization of Heat Shock Transcription Factor 1 in Colorectal Cancer Cells

摘要

摘要:
目的探讨FBXW7对结直肠癌细胞中热休克转录因子1（HSF1）表达和定位的影响。
方法 Western blot法检测敲除FBXW7及WT的结直肠癌细胞系HCT116和DLD1中HSF1及pHSF1^Ser326蛋白的表达；免疫荧光和Western blot检测在热休克及恢复期细胞中pHSF1^Ser326的核蛋白表达和定位。
结果过表达FBXW7α的DLD1细胞中HSF1蛋白表达显著降低（P < 0.01）；敲除FBXW7的HCT116和DLD1细胞中HSF1和pHSF1^Ser326总蛋白表达显著升高（均P < 0.05）。与WT组比较，敲除FBXW7的细胞中HSF1和pHSF1^Ser326主要累积在细胞核，而胞质表达较弱；温热刺激后，WT细胞中HSF1及pHSF1^Ser326表达恢复至未刺激水平，而敲除FBXW7的细胞中HSF1及pHSF1^Ser326在核中有较强表达（均P < 0.01）。
结论敲除FBXW7的结直肠癌细胞热刺激后，细胞核HSF1水平恢复受阻，可能与缺失FBXW7不能降解胞核HSF1有关。

Abstract:
Objective To investigate the effect of FBXW7 on the expression and localization of HSF1 in colorectal cancer cells.
Methods The expression levels of HSF1 and pHSF1^Ser326 protein in FBXW7 deletion (KO) and wild-type (WT) FBXW7-expressing counterpart colorectal cancer cells were detected by Western blot. The nucleoprotein expression and localization of pHSF1^Ser326 in heat-shocked or recovery stage cells were observed by Western blot and immunofluorescence method.
Results The HSF1 expression level in DLD1 cells transfected with FBXW7α was decreased significantly (P < 0.01). The expression levels of HSF1 and pHSF1^Ser326 protein in FBXW7 KO cells were higher than those in WT cells (all P < 0.05). HSF1 and pHSF1^Ser326 in FBXW7 KO cells were mainly expressed in nucleus and weakly expressed in cytoplasm. After warm stimulation, the expression of HSF1 and pHSF1^Ser326 in WT cells recovered to the unstimulated level, while the expression of HSF1 and pHSF1^Ser326 in FBXW7 KO cells were higher in the nucleus (all P < 0.01).
Conclusion Loss of FBXW7 could affect the nuclear HSF1 recovery after warm stimulation. It may be associated with FBXW7 deletion inhibiting the degradation of nuclear HSF1.

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