Effects of shRNA-mediated silence of metastasis-associated lung adenocarcinoma transcript 1 on the invasion and metastasis of ovarian carcinoma cell line OVCAR3

Abstract

Objective To evaluate the expression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1) in ovarian cancer cell lines, and to investigate the biological effects of down-regulated MALAT-1 on OVCAR3 cells. Methods qRT-PCR analysis was used to examine the expression level of MALAT-1 gene in ovarian cancer cells, including ES-2, A2780, SKOV3 and OVCAR3 cell lines. For functional research, four shRNA oligos specially targeting MALAT-1 and a empty vector were designed and constructed into pGPU6/GFP/Neo, then transfected into OVCAR3 cells. qRT-PCR was used to confirm the effective suppression of MALAT-1. Changes of proliferation and adhesion of cells were analyzed by CCK-8 and adhesion assays. Wound-healing, transwell migration and invasion assays were used to examine migration and invasion of MALAT-1-silencing cells in vitro. Results The expression of MALAT-1 gene in OVCAR3 cells was high, and qRT-PCR results confirmed successfully the knockdown of MALAT-1 after transient transfection. After successful suppression of MALAT-1, the proliferation, wound-healing and adhesion ability in vitro were inhibited to some degree. In transwell migration assay, the number of migration cells in MALAT-1-silencing group was 52.17 ± 4.48, which is much less than that in the negative and control groups (286.50 ± 12.23 and 295.67±6.96, respectively). In invasion assay, the number of invasion cells passing the transwell membrane in MALAT-1-silencing group (37.33±2.40) was also decreased significantly, compared to that in the negative and control groups (239.00±15.72 and 222.67±20.85, P< 0.05). Conclusions shRNA-mediated silence of MALAT-1 can effectively inhibit the proliferation, adhesion, migration and invasion abilities of ovarian cancer cell line OVCAR3 in vitro, indicating MALAT-1 is expected to be a target gene for the treatment of ovarian cancer. Key words: Ovarian neoplasms; Metastasis-associated lung adenocarcinoma transcript 1; Neoplasm invasiveness; Neoplasm metastasis; RNA interference