LncRNA-MSC-AS1 inhibits the ovarian cancer progression by targeting miR-425-5p

Abstract

Background: Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) were reported to be aberrantly expressed and related to the pathogenesis of ovarian cancer. However, the role and regulatory mechanism of MSC-AS1 in ovarian cancer has yet to be fully elucidated.

Methods: Expression of lncRNA MSC-AS1 (MSC-AS1) and microRNA-425-5p (miR-425-5p) in the ovarian cancer tissue samples and cell lines was examined by quantitative real-time polymerase chain reaction (qRT-PCR). The functions of MSC-AS1 on ovarian cancer cell proliferation, cell cycle and apoptosis were determined using MTT, colony formation and flow cytometry analyses. The protein expression levels were evaluated using western blot assay. The targeting relationship MSC-AS1 and miR-425-5p was verified via dual-luciferase reporter assay.

Results: MSC-AS1 expression level was lowly expressed, while miR-425-5p level was highly in ovarian cancer tissues and cells. Elevation of MSC-AS1 has the ability to significantly inhibit cell proliferation and facilitate cell apoptosis in SKOV3 and A2780 cells. Moreover, MSC-AS1 targeted and negatively modulated miR-425-5p. MiR-425-5p up-regulation has been proved to partially reverse the tumor suppressive function of MSC-AS1 overexpression CONCLUSION: MSC-AS1 sponged miR-425-5p to inhibit the ovarian cancer progression. These findings may provide a promising therapeutic target for the treatment of ovarian cancer.

Keywords: Apoptosis; LncRNA MSC-AS1; MicroRNA-425-5p; Ovarian cancer; Proliferation.

Fig. 1

MSC-AS1 expression was reduced in ovarian cancer tissues and cell lines. A, B The expression of MSC-AS1 in ovarian cancer tissues and adjacent tissues was predicted using bioinformatics analysis based on GEPIA database. C, D qRT-PCR was used to detect the expression level of MSC-AS1 in ovarian cancer tissues (n=20) and cell lines. ^**P<0.01 vs. Normal tissue or IOSE80

Fig. 2

MSC-AS1 over-expression promotes proliferation of ovarian cancer cells. A The upregulation efficacies of MSC-AS1 in SKOV3 and A2780 cells were validated through qRT-PCR. B, C The impact of MSC-AS1 on cell viability and proliferation was explored by MTT and colony formation analyses. D Flow cytometry analysis was performed to explore the effects of MSC-AS1 on the cell cycle. E The protein expression levels of PCNA and Ki-67 were analyzed using western blot assay. ^*P<0.05, ^**P<0.01 vs. pcDNA 3.1

Fig. 3

Impact of MSC-AS1 on ovarian cancer cell apoptosis. A The apoptosis property of SKOV3 and A2780 cells was investigated by tunel assay. B The apoptosis related protein expressions were investigated via western blot assay. ^*P<0.05, ^**P<0.01 vs. pcDNA 3.1

Fig. 4

MiR-425-5p is a direct target of MSC-AS1. A The binding relationship between MSC-AS1 and miR-425-5p was assumed by StarBase datebase. B Level of miR-425-5p in ovarian cancer tissues and normal tissues was evaluated by qRT-PCR assay (n=20). C miR-425-5p expression was detected in ovarian cancer cell lines. D Luciferase reporter assay assessed the target of MSC-AS1 to miR-425-5p. E qRT-PCR analysis was used to measure the expression of miR-425-5p under transfection of pcDNA 3.1-MSC-AS1 or pcDNA 3.1. ^**P<0.01 vs. Normal tissues, IOSE80, or pcDNA 3.1

Fig. 5

MSC-AS1 inhibits the ovarian cancer progression by targeting miR-425-5p. A, B Transfection efficiency was analyzed. C, D Cell viability and proliferation was measured by using MTT and EdU assays. ^**P<0.01, ^***P<0.001 vs. pcDNA 3.1. ^##P<0.01 vs. pcDNA 3.1 MSC-AS1

Fig. 6

MSC-AS1 inhibits the ovarian cancer progression by targeting miR-425-5p. A, B. Determination of cell cycle and apoptosis by flow cytometry analysis. ^*P<0.05, ^**P<0.01 vs. pcDNA 3.1. ^#P<0.05, ^##P<0.01 vs. pcDNA 3.1 MSC-AS1