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. 2019 Jul 10:24:49.
doi: 10.1186/s11658-019-0172-y. eCollection 2019.

Silencing of Synuclein-γ inhibits human cervical cancer through the AKT signaling pathway

Chunnian Zhang  1 , Liqin Gu  1 , Xiafang Li  1 , Jianzhong Wang  1
Affiliations

Silencing of Synuclein-γ inhibits human cervical cancer through the AKT signaling pathway

Chunnian Zhang et al. Cell Mol Biol Lett. .

Abstract

Background: Synuclein-γ has been demonstrated to be highly expressed in various human cancers including cervical cancer, and has been shown to play a critical role in tumor aggressiveness. We aimed to investigate the role of Synuclein-γ in human cervical cancer in vitro and in vivo.

Method: Reverse transcription-quantitative polymerase chain reaction assay and Western blot assay were used to detect the mRNA and protein expression, respectively. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and colony formation assay were performed to measure the viabilities of cancer cells. Flow cytometry assay was used to detect the cell cycle and apoptosis. Moreover, an animal experiment was performed to evaluate the biological behavior of Synuclein-γ in vivo.

Results: In the current study, we found that Synuclein-γ was obviously over-expressed in cervical cancer tissues compared to the adjacent non-cancer tissues. Cervical cancer cells transfected with Synuclein-γ siRNA demonstrated significant inhibition of cancer proliferation (P < 0.01), cell cycle arrest at G0/G1 phase, and cell apoptosis (P < 0.05). Moreover, down-regulation of Synuclein-γ significantly inhibited cervical cancer growth in vivo. In addition, protein levels of AKT, c-Myc and Cyclin D1 were much lower in the Synuclein-γ siRNA-treated groups than that in the control group.

Conclusions: Synuclein-γ inhibition reduced cervical cancer tumor growth through the AKT pathway. This effect represented a therapeutic opportunity and provided a novel target for cervical cancer treatment.

Keywords: AKT; Cervical cancer; Growth; Knockdown; Synuclein-γ.

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Conflict of interest statement

Competing interestsThe authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
SNCG gene was up-regulated in cervical cancer. a Total RNAs were isolated from human cervical cancer tissues and adjacent non-tumor tissues. qRT-PCR was performed to determine the SNCG expression in human samples. b Protein expression level of SNCG in human cervical cancer tissues and adjacent non-tumor tissues. c qRT-PCR was performed to determine the SNCG expression in human cervical epithelial (HCerEpiC) cells in two human cervical cancer cell lines (HeLa, SiHa). d Protein expression levels of SNCG in human cervical cancer cell lines were detected. * P < 0.05, ** P < 0.01,*** P < 0.001
Fig. 2
Fig. 2
Silencing of SNCG inhibited proliferation and transformation of cervical cancer cells a SNCG-RNAi down-regulated SNCG mRNA expression in HeLa and SiHa cells. Total RNA was extracted and qRT-PCR was performed. b Effect of SNCG siRNA on cell growth determined by MTT assay. c Colony formation of HeLa and SiHa cells transfected with SNCG siRNA. * P < 0.05, ** P < 0.01,*** P < 0.001
Fig. 3
Fig. 3
Silencing of SNCG arrested HeLa cells in the G0/G1 phase and induced apoptosis (a) The proportion of cells in G0/G1 phase significantly increased while the proportion in G2/M phase decreased in the SNCG siRNA group. b Detection of apoptosis 48 h after transfection with siSNCG using FCM analysis. SNCG RNAi significantly promoted cell apoptosis in the SNCG siRNA group. ** P < 0.01
Fig. 4
Fig. 4
Silencing of SNCG inhibited the growth of cervical cancer cells in vivo a Tumors extracted at 27 days. b Weight curves of tumors at 27 days. Tumor weights were significantly decreased after SNCG siRNA. (***P < 0.001)
Fig. 5
Fig. 5
Silencing of the SNCG gene affects the AKT signaling pathways. a Western blotting of phosphorylated AKT (p-Akt), c-Myc, Cyclin D1 and their corresponding internal reference (β-actin). The levels of p-AKT, c-Myc, Cyclin D1 were lower in the SNCG siRNA group than NC and CON groups while there was no significant difference in expression of β-actin between the two groups. b Effect of SNCG on cell growth determined by MTT assay. LY294002 significantly reversed the upregulatory effect of SNCG on cell proliferation
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