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Archive > Volume 24 Issue 12 > 2017,24(12):1391-1396. DOI:10.3872/j.issn.1007-385X.2017.12.009 Prev Next
Basic Research
Roles of zinc finger protein 148 alternative splicing isoforms in occurrence and development of colorectal cancer
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Abstract:
Objective：To investigate the effect of two alternative splicing isoforms of zinc finger protein 148 (ZNF148) gene on the invasion and metastasis of colon cancer cells and their related mechanisms. Methods: RTPCR was used to determine the expressions of two ZNF148 alternative splicing isoforms in the human colorectal SW480 cells. ZNF148 interference vector and ZNF148 over-expression vector were constructed and transfected into the SW480 cells; them were divided into ZNF148FL-siRNA group, ZNF148FL-Over express group, ZNF148ΔN-siRNA group, ZNF148ΔN- Over express group, Control siRNA group, Control Over express group and Normal control group. The mRNA expressions in each group were examined by RT-PCR; the proliferation, invasion and migration in vitro as well as apoptosis of SW480 cells were detected by MTT, Transwell, scratch assay and flow cytometry, respectively.Results: Two splicing isoforms (ZNF148FL of 2 385 bp and ZNF148ΔN of 2 004 bp) were obtained by RTPCR.The expression level of ZNF148FL was significantly decreased while the expression of ZNF148ΔN was increased in the ZNF148FL-siRNA group; the expression of ZNF148FL was significantly increased while the expression of ZNF148ΔN was significantly decreased in the ZNF148FL- Over express group (all P<0.050. The expression of ZNF148ΔN was significantly decreased while the expression of ZNF148FL was increased in ZNF148ΔN-siRNA group;the expression of ZNF148ΔN was significantly increased while the expression of ZNF148FL was decreased in ZNF148ΔN-over express group (all P<0.05). The proliferation of the SW480 cells was increased in ZNF148FL-over express group and the ZNF148ΔN- siRNA group, while the proliferation of the SW480 cells was decreased in the ZNF148FL-siRNA group and the ZNF148ΔN-Over express group. The transmembrane cell number and migration ability of the SW480 cells in the ZNF148FL-siRNA group and the ZNF148ΔN-Over express group were significantly decreased,but the apoptotic rate was significantly increased; However, ZNF148FL-Over Express group and ZNF148ΔNsiRNA group showed the significantly increased transmembrane cell number and migration ability but decreased apoptosis rate (all P<0.05). Conclusion: ZNF148FL could increase proliferation, invasion and metastasis of colorectal cancer cells, while ZNFΔN showed opposite effect; the two splicing isoforms of ZNF148 may exert mutual antagonistic effect to each other on the malignant biological activities.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No. 81572332）
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