Effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells / 西安交通大学学报(医学版)

ABSTRACT

【Objective】 To investigate the effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells and its mechanism. 【Methods】 Human gastric cancer cells MKN-45， AGS and gastric mucosal epithelial cells GES-1 were cultured； circ_0045943 was detected by real-time polymerase chain reaction. The overexpression and silencing of circ_0045943 adenovirus vectors OE-circRNA and sh-circRNA together with their negative controls OE-NC and sh-NC were constructed and transfected； CCK-8 method was used to detect the proliferation activity of AGS cells after overexpression and silencing of circ_0045943； TUNEL method was used to detect the cell apoptosis； transwell assay was used to detect the cell migration and invasion； and would healing assay was used to detect the cell migration. Starbase database screened the binding site of miR-106a and circ_0045943. Real-time PCR was used to detect the expression of miR-106a， and the expression of circ_0045943 and the changes of miR-106a after the treatment of OE-circRNA and sh-circRNA. 【Results】 Real-time PCR showed that the expression of circ_0045943 decreased in gastric cancer cells MKN-45 and AGS compared to GES-1 （Pboth<0.001）. CCK-8 showed that the absorbance value of AGS cells in OE-circRNA group was lower than that in sh-circRNA group （P24 h<0.01， P48 h<0.001， and P72 h<0.001）. TUNEL showed the number of apoptotic AGS cells increased after overexpression of circ_0045943， but decreased after silencing of circ_0045943. Transwell assay showed that the migration and invasion of AGS cells were lower in OE-circRNA group than in sh-circRNA group （Pboth<0.001）. The wound healing assay showed that the migration rate of AGS cells in OE-circRNA group was the lowest， but was high in sh-circRNA group （P<0.001）. Starbase retrieved that circ_0045943 and miR-106a had complementary binding sequences. Real-time PCR showed that miR-106a was highly expressed in gastric cancer cells （P<0.001）， and the expressions of circ_0045943 and miR-106a significantly differed after treatment with OE-circRNA and sh-circRNA （Pboth<0.001）. With the increase or decrease of circ_0045943， the expression of miR-106a changed in the opposite direction. 【Conclusion】 Circ_0045943 has low expression in gastric cancer， and promoting or inhibiting circ_0045943 expression may regulate the proliferation， apoptosis， migration and invasion of gastric cancer cells by targeting miR-106a.