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. 2017 Dec;14(6):6519-6524.
doi: 10.3892/ol.2017.7009. Epub 2017 Sep 21.

Knockdown of TGIF attenuates the proliferation and tumorigenicity of EC109 cells and promotes cisplatin-induced apoptosis

Yadong Wang  1   2 , Teng Pan  3 , Li Li  1 , Haiyu Wang  1 , Jiangmin Li  1 , Ding Zhang  1 , Haiyan Yang  3
Affiliations

Knockdown of TGIF attenuates the proliferation and tumorigenicity of EC109 cells and promotes cisplatin-induced apoptosis

Yadong Wang et al. Oncol Lett. 2017 Dec.

Abstract

A previous study has reported that frequent amplifications of the TG-interacting factor (TGIF) were observed in esophageal squamous cell carcinoma. The aim of the present study was to investigate the potential role of TGIF in the proliferation and tumorigenicity of the esophageal cancer cell line EC109 and cisplatin-induced apoptosis. Stable TGIF-knockdown EC109 cell line was established by infecting short hairpin RNA (shRNA) lentiviral particles. Soft agar and tumor xenograft assays were applied in nude mice. Flow cytometry was employed to evaluate the cell cycle and apoptosis. Western blot analysis was used to detect the expression of proteins. TGIF knockdown suppressed EC109 cell proliferation, colony formation in soft agar and tumor growth in nude mice, induced cell cycle arrest in the G1 phase, and promoted cisplatin-induced apoptosis. In addition, TGIF knockdown significantly reduced the expression of phospho-Rb in EC109 cells. The reduced level of full length PARP expression and the increased level of cleaved caspase-3 expression were observed in EC109 cells with the treatment of cisplatin and TGIF knockdown. The results suggest that knockdown of TGIF attenuated the proliferation and tumorigenicity of EC109 cells, and promoted cisplatin-induced apoptosis.

Keywords: TG-interacting factor; apoptosis; cell cycle; esophageal cancer; tumorigenicity.

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Figures

Figure 1.
Figure 1.
(A) The identification of TGIF-knocked down EC109 cells. Our data indicated that shRNA significantly reduced the level of TGIF protein expression in EC109 cells. (B) The effects of TGIF knockdown on EC109 cell proliferation. Our results showed that TGIF knockdown inhibited EC109 cell proliferation from 72 h (*P<0.05 vs. shRNA-control). (C) Effects of TGIF knockdown on colony formation. Our findings indicated that the ability of colony formation was significantly decreased in EC109-shRNA-TGIF cells as compared with EC109-shRNA-control cells. (D) Effects of TGIF knockdown on tumor growth in nude mice. Our results demonstrated that the tumor weight was significantly reduced in EC109-shRNA-TGIF cells as compared with EC109-shRNA-control cells. TGIF, TG-interacting factor; shRNA, short hairpin RNA.
Figure 2.
Figure 2.
(A) Effects of TGIF knockdown on cell cycle distribution. Data showed that TGIF knockdown increased the percentage of EC109 cells in G1 phase and decreased the percentage of EC109 cells in S phase. (B and C) Effects of TGIF knockdown on the expression of interested proteins. Data showed that TGIF knockdown suppressed the expression of phospho-Rb protein. TGIF, TG-interacting factor; shRNA, short hairpin RNA.
Figure 3.
Figure 3.
Effects of cisplatin on the expression of TGIF protein. Data showed that cisplatin could reduce the expression of TGIF protein. TGIF, TG-interacting factor.
Figure 4.
Figure 4.
(A) Effects of TGIF knockdown on cisplatin-induced EC109 cells apoptosis. Data showed that the percentage of total apoptosis was significantly increased in EC109-shRNA-TGIF cells treated with cisplatin compared with EC109-shRNA-control cells treated with cisplatin. (B) Effects of TGIF knockdown on apoptosis-related markers. TGIF knockdown decreased the expression of full length PARP and increased the expression of cleaved caspase-3 in cisplatin-treated group. TGIF, TG-interacting factor; shRNA, short hairpin RNA.
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