Hsa-miR-218 inhibits growth of cervical cancer HeLa cells by targeting to LASP1 |
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Authors: | QIU Yu HUANG Jian-ping ZHOU Qin-xian WANG Huan SHI Hui-gang HE Jin-hua |
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Affiliation: | 1. Department of Obstetrics & Gynaecology, Ji'an Maternal and Child Health-Care Hospital, Ji'an 343000, China;2. Nangji Biochemistry Technical Limited Company of Guangzhou, Guangzhou 510360, China;3. Department of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou 511400, China |
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Abstract: | AIM: To study the effect of hsa-miR-218 on cervical cancer HeLa cell growth and the underlying molecular mechanism.METHODS: The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed. pmiR-218 was transfected into HeLa cells. The number of viable HeLa cells was counted by the method of Trypan blue exclusion. The inhibitory rate of cell activity was detected by WST-8 assay. The expression of LIM and SH3 protein 1(LASP1) at mRNA and protein levels was determined by real-time PCR and Western blot. The interaction between miR-218 and LASP1 was examined using a luciferase reporter assay.RESULTS: The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed successfully and confirmed by DNA sequencing. Over-expression of miR-218 inhibited the activity of HeLa cells with the inhibitory rates of 15%, 26% and 65% at 24 h, 48 h and 72 h, respectively. The difference between transfection group and blank control/negative control group was statistically significant. The luciferase activity was reduced when co-transfection with miR-218 mimics and LASP1-3,UTR plasmid. The relative expression of miR-218 was increased after transfection with pmiR-218. Over-expression of miR-218 down-regulated the LASP1 expression at mRNA and protein levels by 25% and 75% respectively. Compared with blank control group and negative control group, the difference was statistically significant(P<0.05).CONCLUSION: pmiR-218 effectively inhibits the growth of HeLa cells in a time-dependent manner. miR-218 targets to the 3,UTR of LASP1, thus down-regulating the expression of LASP1 in HeLa cells. |
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Keywords: | Hsa-miR-218 LIM and SH3 protein 1 Cervical cancer HeLa cells |
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