Effect of the miR‑96‑5p inhibitor and mimic on the migration and invasion of the SW480‑7 colorectal cancer cell line

Abstract

The objectives of the present study were to identify the aberrant expression of microRNA (miRNA) in colorectal carcinoma (CRC) tissues from published miRNA profiling studies and to investigate the effects of the identified miRNA inhibitor and mimic miR-96-5p on CRC cell migration and invasion. The altered expression of the regulators of cytoskeleton mRNA in miR-96-5p inhibitor-transfected cells was determined. The miR-96-5p expression level in five CRC cell lines, HCT11, CaCo2, HT29, SW480 and SW620, and 26 archived paraffin-embedded CRC tissues were also investigated by reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR). Cell viability in response to the miR-96-5p inhibitor and mimic transfections was determined by an MTT assay. A Matrigel invasion assay was conducted to select the invasive subpopulation designated SW480-7, by using the parental cell line SW480. The effects of miR-96-5p mimic-or inhibitor-transfected SW480-7 cells on cell migration and invasion were evaluated using the Transwell and Matrigel assays, and the change in expression of the regulators of cytoskeleton mRNAs was identified by Cytoskeleton Regulators RT 2-Profiler PCR array followed by validation with RT-qPCR. CRC tissues exhibited a significant increase in miR-96-5p expression, compared with their matched normal adjacent tissues, indicating an oncogenic role for miR-96-5p. The results demonstrated that the miR-96-5p inhibitor decreased the migration of SW480-7 cells, but had no effect on invasion. This may be due to the promotion of cell invasion by Matrigel, which counteracts the blockade of cell invasion by the miR-96-5p inhibitor. The miR-96-5p mimic enhanced SW480-7 cell migration and invasion, as expected. It was determined that there was a >2.5 fold increase in the expression of genes involved in cytoskeleton regulation, myosin light chain kinase 2, pleckstrin homology like domain family B member 2, cyclin A1, IQ motif containing GTPase activating protein 2, Brain-specific angiogenesisinhibitor 1-associated protein 2 and microtubule-actin crosslinking factor 1, in miR-96-5p inhibitor-transfected cells, indicating that they are negative regulators of cell migration. In conclusion, the miR-96-5p inhibitor blocked cell migration but not invasion, and the latter may be due to the counteraction of Matrigel, which has been demonstrated to stimulate cell invasion.

microRNA-96 promotes occurrence and progression of colorectal cancer via regulation of the AMPKα2-FTO-m6A/MYC axis

“…This value may be correlated with the role of miR-96 [27]. More recently, miR-96 inhibitor could function as a suppressor of cell migration instead of cell invasion that may be affected by the counteraction of cell invasion stimulator Matrigel [13]. In the present study, we validated the anti-tumor activity exerted by miR-96 antagomir in the nude mouse model.…”

“…Interestingly, miR-96 has been widely implicated in the progression and metastatic potential of a plethora of cancers, including cervical [11], and ovarian [12] cancers. Although miR-96 is witnessed to link to tumor invasion in addition to cancer cell migration and invasion in CRC [7,13], little is well-understood regarding its downstream mechanisms related to the pathophysiological events of CRC.…”

An in silico approach to identify and prioritize miRNAs target sites polymorphisms in colorectal cancer and obesity

“…The miRNAs in prioritized interactions were related to CRC and other type of cancers. In this case, the miR‐30 family, miR‐504, miR‐34c, miR‐488, miR‐15b, miR‐195, miR‐497, miR‐15a, miR‐503, miR‐141, miR‐182, miR‐96, and miR‐136 were also related to invasion, proliferation, metastasis and tumor growth, or survival of several cancers including CRC 46‐65 . However, there are few studies on the role of included prioritized miRNAs for interactions with obesity.…”

“…In this case, the miR-30 family, miR-504, miR-34c, miR-488, miR-15b, miR-195, miR-497, miR-15a, miR-503, miR-141, miR-182, miR-96, and miR-136 were also related to invasion, proliferation, metastasis and tumor growth, or survival of several cancers including CRC. [46][47][48][49][50][51][52][53][54][55][56][57][58][59][60][61][62][63][64][65] However, there are few studies on the role of included prioritized miRNAs for interactions with obesity. For obesity there are some studies on mice, for instance, miR-16-5p decrease with high weight and a mutation in miR-16 gene lead to increasing in body weight.…”

Antigens from H22 cell mutation induced by lentinan are correlated with HCC immunoprophylaxis

“…Among them, cellular immunity plays a leading role, and humoral immunity and cellular immunity play an antitumor role [28]. [34]. Therefore, the inhibition of angiogenesis is an effective means to inhibit the growth and metastasis of tumor cells.…”