Inhibition of cell migration and invasion by miR‑29a‑3p in a colorectal cancer cell line through suppression of CDC42BPA mRNA expression

Abstract

The objective of this study was to determine the effect of miR‑29a‑3p inhibitor on the migration and invasion of colorectal cancer cell lines (CRC) and the underlying molecular mechanisms. miR‑29a‑3p was detected using reverse transcription-quantitative polymerase chain reaction (RT‑qPCR) in the CRC cell lines HCT11, CaCo2, HT29, SW480 and SW620. An invasive subpopulation designated SW480‑7 was derived from the parental cell line, detected by Transwell and Transwell Matrigel assays. Cytoskeleton Regulators RT2 profiler PCR array and western blot analysis were utilized to identify the alterations in expression of downstream mRNAs. siRNA against CDC42BPA was transfected into SW480‑7 and effects on cell migration and invasion were investigated. Data obtained showed that miR‑29a‑3p was detected in these five CRC cell lines. miR‑29a‑3p inhibitor had no effect on viability but stimulated cell migration and invasion of SW480‑7 cells. In contrast, miR‑29a‑3p mimic suppressed cell migration and invasion. TargetScan miRBD and DIANA were employed to identify the potential direct target genes of miR‑29a‑3p in the Cytoskeleton Regulators RT2-Profiler PCR array. Cytoskeleton Regulators RT2-Profiler PCR array data showed that 3 out of the 5 predicted targets genes, CDC42BPA (2.33-fold), BAIAP2 (1.79-fold) and TIAM1 (1.77-fold), in the array were upregulated by miR‑29a‑3p. A significant increase in expression IQGAP2, PHLDB2, SSH1 mRNAs and downregulation of PAK1 mRNA was also detected with miR‑29a‑3p inhibition. Increase in CDC42BPA, SSH1 and IQGAP2 mRNA expression correlated with increased protein level in miR‑29a‑3p transfected SW-480-7 cells. Silencing of CDC42BPA (an enhancer of cell motility) partially abolished miR‑29a‑3p inhibitor-induced stimulation of cell migration and invasion. miR‑29a‑3p expression in stage II and III CRC is relatively lower than that of stage I CRC. However, the data need to be interpreted with caution due to the small sample size. In conclusion, inhibition of miR‑29a‑3p stimulates SW480‑7 cell migration and invasion and downstream expression IQGAP2, PHLDB2, SSH1 mRNAs are upregulated whilst PAK1 mRNA is downregulated. Silencing of CDC42BPA expression partially reduces miR29a‑3p inhibitor-induced migration and invasion of SW480‑7 cells.

LncRNA testis-specific transcript, Y-linked 15 (TTTY15) promotes proliferation, migration and invasion of colorectal cancer cells via regulating miR-29a-3p/DVL3 axis

“…It should be noted that the role of miR-29a-3p in CRC has also been reported. Down-regulation of miR-29a-3p can promote the invasion and migration of CRC cells, and it can repress the invasion and migration of cancer cells by reducing the expression of CDC42BPA [16]. We observed that in CRC tissues, the expression of TTTY15 and miR-29a-3p was notably negatively correlated.…”

“…The expression of miR-29a-3p is also significantly down-regulated in gastric cancer and its low expression facilitate the proliferation, migration and invasion of cancer cells [14]. There are also studies on the expression and mechanism of miR-29a-3p in CRC [15,16], but the research is still not perfect, and its upstream and downstream molecular mechanisms still need to be further studied.…”

miR-29a-5p Regulates the Proliferation, Invasion, and Migration of Gliomas by Targeting DHRS4

“…The above results show us the two sides of mi-29a. A further functional study has indicated that miR−29a−3p inhibits cell migration and invasion of a colorectal cancer cell line by suppressing CDC42BPA mRNA expression (30). In our other experiment, through gene sequencing, it was found that miR-29a-5p was highly expressed in neural stem cells and inhibited its differentiation into glial cells.…”

P21-Activated Kinase 1: Emerging biological functions and potential therapeutic targets in Cancer

“…In addition, PAK1 can be directly activated by interaction with Rho-related GTPases CDC42 and RAC1 86 . However, some miRNAs targeting these activators can also regulate the activity of PAK1, such as miR-142-3p and miR-4715-5p targeting RAC1 87 , 88 , and miR302-367, miR‑29a‑3p, and miR-15b targeting CDC42 48 , 89 , 90 . Other miRNAs perform other functions.…”