Effect of inhibition to Yes-related proteins-mediated Wnt/β-catenin signaling pathway through miR-195-5p on apoptosis of gastric cancer cells

D.-L. Zhao, Q.-L. Wu

Department of Gastrointestinal Surgery, Zhuji People’s Hospital, Shaoxing, P.R. China. dsjv28@163.com

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• Oncology

OBJECTIVE: This clinical study aimed to explore the expression of miR-195-5p and Yes-associated protein (YAP) in gastric cancer (GC) and their relevant apoptosis mechanism.
PATIENTS AND METHODS: Thirty GC patients treated in our hospital were enrolled as a patient group and 30 normal people who underwent physical examination during the same period were enrolled as a normal group. The purchased GC cells and human gastric mucous membrane cells were used to transfect miR-195-5p-mimics, miR-NC, si-YAP, and si-RNA into MKN45 and SGC7901 cells. qRT-PCR was used to detect the expression of miR-195-5p and YAP in samples, and WB was used to detect the expression of YAP, Caspase-3, Caspase-9, β-catenin, c-myc, and cyclin Dl. CCK-8, the transwell, and the flow cytometry were used to detect cell proliferation, invasion, and apoptosis. The dual fluorescent enzyme reporter was used to determine the relationship between miR-195-5p and YAP.
RESULTS: MiR-195-5p was lowly expressed in the tissues and serum of patients, but YAP was highly expressed, and the area under the miR-195-5p and YAP curves was more than 0.9. MiR-195-5p and YAP were associated with tumor diameter, TNM stage, lymph node metastasis, and differentiation in GC patients. The overexpression of miR-195-5p and the inhibition of YAP expression can inhibit cell proliferation and invasion and promote apoptosis. WB assay showed that the overexpression of miR-195-5p and the inhibition of YAP could inhibit the expression of β-catenin, c-myc, and cyclin D1 protein and promote the expression of Caspase-3, Caspase-9 protein. The dual fluorescent enzyme reporter revealed that there was a targeting relationship between miR-195-5p and YAP.
CONCLUSIONS: The overexpression of miR-195-5p can inhibit YAP-mediated Wnt/β-catenin signaling pathway and promote cell apoptosis, so it may be a potential therapeutic target for GC.

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