Effects of shRNA-mediated silence of metastasis-associated lung adenocarcinoma transcript 1 on the invasion and metastasis of ovarian carcinoma cell line OVCAR3
10.3760/cma.j.issn.1006-9801.2015.10.004
- VernacularTitle:shRNA介导沉默肺腺癌转移相关转录本1表达对人卵巢癌细胞株OVCAR3侵袭与转移的影响
- Author:
Yanqing ZHOU
;
Juan LI
;
Linyu TAN
;
Xiaying XU
;
Huabing LYU
;
Qirong WEN
;
Xiujie SHENG
- Publication Type:Journal Article
- Keywords:
Ovarian neoplasms;
Metastasis-associated lung adenocarcinoma transcript 1;
Neoplasm invasiveness;
Neoplasm metastasis;
RNA interference
- From:
Cancer Research and Clinic
2015;27(10):664-668,672
- CountryChina
- Language:Chinese
-
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-l-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.
