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. 2016 May;11(5):3123-3130.
doi: 10.3892/ol.2016.4345. Epub 2016 Mar 17.

Downregulation of SASH1 correlates with tumor progression and poor prognosis in ovarian carcinoma

Xiaoyan Ren  1 , Yifei Liu  2 , Yumei Tao  1 , Guoxiang Zhu  1 , Meilan Pei  3 , Jianguo Zhang  2 , Jian Liu  4
Affiliations

Downregulation of SASH1 correlates with tumor progression and poor prognosis in ovarian carcinoma

Xiaoyan Ren et al. Oncol Lett. 2016 May.

Abstract

SAM- and SH3-domain containing 1 (SASH1) is a recently identified tumor suppressor gene that is required in the tumorigenesis of breast and other solid carcinomas. The SASH1 protein contains SH3 and SAM domains, indicating that it may serve an important role in intracellular signal transduction. The purpose of the present study was to investigate the expression of SASH1 in ovarian carcinoma and the correlation between its expression with clinical pathological features and clinical significance, and the effect of SASH1 on cell proliferation, apoptosis and migration of ovarian SKOV3 cells. The human ovarian carcinoma tissues and adjacent normal tissues were collected following surgery. Reverse transcription-quantitative polymerase chain reaction and western blot analysis were used to detect the expression levels of SASH1 mRNA and protein, respectively. The expression levels of SASH1 mRNA and protein in ovarian carcinoma tissues were significantly lower than that observed in adjacent normal tissues (P<0.05). The expression levels of SASH1 in samples from patients without lymph nodes metastasis and patients with early FIGO stage was lower than those with lymph nodes metastasis and patients with advanced FIGO stage (P<0.05). Flow cytometry analysis and Transwell invasion chamber experiments were used to investigate the effect of SASH1 on the cell proliferation, apoptosis and migration of SKOV3 cells. The recombinant plasmid pcDNA3.1-SASH1 was constructed and transfected into SKOV3 cells. In addition, the SKOV3 cells in the pcDNA3.1-SASH1 group exhibited significantly reduced cell growth, proliferation, and migration ability compared to the empty vector group and normal group (P<0.01). There were a greater number of apoptotic cells in the pcDNA3.1-SASH1 group compared to the empty vector group and normal group (P<0.01). Taken together, these results indicated that SASH1 may be a tumor suppressor gene in ovarian carcinoma, and SASH1 expression inhibited growth, proliferation and migration, and enhanced apoptosis of SKOV3 cells.

Keywords: SASH1; apoptosis; migration; ovarian cancer; proliferation.

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Figures

Figure 1.
Figure 1.
SASH1 mRNA expression levels were analyzed in ovarian cancer tissues by reverse transcription-quantitative polymerase chain reaction. (A) Normal surrounding ovarian tissues; (B) ovarian cancer tissues; (C) negative lymph node metastasis; (D) positive lymph node metastasis; (E) patients in FIGO stages (I, II); (F) patients in FIGO stages (III, IV). *P<0.05 was considered to indicate a significant difference. SASH1, SAM- and SH3-domain containing 1; FIGO, International Federation of Gynaecologists and Obstetricians.
Figure 2.
Figure 2.
SASH1 protein expression was analyzed in ovarian cancer tissues by western blotting. (A) Relatively normal surrounding ovarian tissues; (B) ovarian cancer tissues; (C) negative lymph node metastasis; (D) positive lymph node metastasis; (E) patients in FIGO stages (I, II); (F) patients in FIGO stages (III, IV). *P<0.05 was considered to indicate a significant difference. SASH1, SAM- and SH3-domain containing 1; FIGO, International Federation of Gynaecologists and Obstetricians.
Figure 3.
Figure 3.
The SASH1 protein expression in SKOV3 cells was assessed by western blotting after transfection. **P<0.01 vs. normal (untransfected) control group. SASH1, SAM- and SH3-domain containing 1.
Figure 4.
Figure 4.
The effect of SASH1 on SKOV3 cell proliferation was analyzed by flow cytometry. **P<0.01 vs. normal (untransfected) control group. SASH1, SAM- and SH3-domain containing 1.
Figure 5.
Figure 5.
Effect of SASH1 on SKOV3 cell growth. The cell growth curve of SKOV3 cells that were transfected with recombined vector pcDNA3.1-SASH1 or empty vector pcDNA3.1. The cell number was assessed by microscopically cell counting after Hoechst 33342 dye staining. *P<0.05 vs. normal (untransfected) control group. SASH1, SAM- and SH3-domain containing 1
Figure 6.
Figure 6.
The effect of SASH1 on SKOV3 cells apoptosis was analyzed by flow cytometry. **P<0.01 vs. normal (untransfected) control group. SASH1, SAM- and SH3-domain containing 1.
Figure 7.
Figure 7.
The effect of SASH1 on SKOV3 cells invasion was analyzed by Transwell invasion chamber. The cell number that passed through the polycarbonate membrane was assessed by microscopically cell counting after Hoechst 33342 staining. **P<0.01 vs. normal (untransfected) control group. SASH1, SAM- and SH3-domain containing 1
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