ABSTRACT
BACKGROUND: Circular RNAs (circRNAs) are involved in the progression of colon cancer (CC). This study aimed to examine the role of a new circRNA circ_0101050 in CC. METHODS: Dual-luciferase reporter and RNA immunoprecipitation analyses were performed to validate the target relationships among maternal embryonic leucine zipper kinase (MELK), microRNA (miR)-140-3 p, and circ_0101050. Expression levels were calculated using western blotting and/or quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Western blotting was performed to evaluate the relative expression of Bcl-2 and Bax proteins to determine cell death. Cell Counting Kit-8 (CCK-8) and colony formation assays were performed to determine the proliferative potential of CC cells. The migration rate of CC cells was evaluated using wound healing assays. Tumor formation tests were performed to determine the effect of circ_0101050 on tumor development in vivo. RESULTS: Elevated levels of circ_0101050 and MELK were observed in CC. By inhibiting circ 0,101,050 or MELK, CC cell proliferation and migration were inhibited, but CC cell apoptosis was promoted. Silencing circ_0101050 also inhibited CC growth in vivo. We also found that miR-140-3 p was downregulated, which alleviated the repressive effects of circ_0101050 knockdown on proliferating and migrating CC cells, as well as the stimulating effect on apoptosis. In addition, the absence of MELK alleviated the effects of miR-140-3 p downregulation, which enhanced CC cell malignancy. CONCLUSIONS: Circ_0101050 exacerbates malignant phenotypes in CC by targeting the miR-140-3 p/MELK axis. These findings suggested that the circ_0101050/miR-140-3 p/MELK network may be a prospective target for CC treatment.
ABSTRACT
OBJECTIVE: To survey the role of hepatocyte growth factor activator inhibitor type 1 (HAI-1) in early embryo development, we study the HA-1 gene expressing in villous tissue after embryo implantation. METHODS: Fifty villous tissue samples from pregnant 6 w to 10 w were divided into five groups and each group included ten samples. The HAI-1 gene and protein expressing in the villous tissue were detected by RT-PCR and immunohistochemistry. RESULTS: The expression of HAI-1 had a gradual increase with the pregnancy weeks added. The HAl-1 gene expression was very low at pregnant 6 w(A(260/280) 0.3142), From pregnant 7 w to 9 w, the expression was increased gradually (A(260/280) 0.3476 in 7 w and 0.3910 in 8 w) and reached the pinnacle in 9 w (A(260/280) 0.4876). And then the pinnacle value lasted until 10 w (A(260/280) 0.4909). The proportion of HAI-1 mRNA/beta-actin mRNA between 9 w and 10 w was not significantly different (P > or = 0.05). But among the other 3 groups, the proportion was significantly different (P < 0.01). The proportion of 9 w and 10 w was significantly different from the other 3 groups (P < 0.01). The results of HAI-1 protein expression showed the similar conclusion by immunohistochemical analysis. CONCLUSION: The HAI-1 gene may play an important role in early embryo development in human.
