Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4
10.3969/j.issn.1007-3969.2015.01.002
- VernacularTitle:下调miR-21对PDCD4表达及结肠癌HT-29细胞功能的影响研究
- Author:
Yupeng REN
;
Chun SONG
;
Hao ZHANG
- Publication Type:Journal Article
- Keywords:
Colon cancer;
Proliferation;
Apoptosis;
miR-21;
HT-29
- From:
China Oncology
2015;(1):6-12
- CountryChina
- Language:Chinese
-
Abstract:
Background and purpose: PDCD4 may be inhibited by miR-21 to regulate the malignant behaviors of colon cancer such as invasion and migration. This study aimed to explore the function of colon cancer HT-29 cell lines by downregulating miR-21 expression and discuss the mechanisms and relationship between miR-21 and PDCD4 in colon cancer malignant behaviors. Methods:simiR-21 was transfected into colon cancer cell line HT-29 to downregulate the expression of miR-21. Proliferation, apoptosis, migration and invasion were detected by MTT, flow cytometry and Transwell assay after transfection. PDCD4 expression was detected by Western blot and qRT-PCR. Results:The qRT-PCR analysis result proved that the transfection efifciency was 60%-65%. MTT analysis result showed that the proliferations of HT-29 cells were inhibited after the transfection of miR-21 for 72, 96, 120 h (t=1.276, P<0.05;t=3.276, P<0.01;t=4.523, P<0.01). Comparing with si-negative control and miR-21 groups, lfow cytometry result showed that the apoptosis rate was increased after miR-21 expression downregulated (t=2.132, P<0.05;t=3.524, P<0.05). Transwell assay result showed that migration (t=2.423, P<0.05; t=3.153, P<0.05) and invasion(t=3.245, P<0.05; t=5.236, P<0.05) were inhibited;Western blot result showed that PDCD4 expression was up-regulated at protein level(t=2.342, P<0.05;t=4.215, P<0.05);qRT-PCR result showed that PDCD4 expression was up-regulated at mRNA level(t=2.261, P<0.05; t=3.492, P<0.05). Conclusion: The proliferation, migration and invasion are the inhibited, and apoptosis is attenuated after miR-21 downregulated by simiR-21 transfection, PDCD4 expression is up-regulated. miR-21 may enhance the malignant behavior of cancer cells by downregulating the PDCD4 expression, miR-21 might be a target gene for colon cancer therapy.
