超声微泡造影剂携带pcDNA-STC1转染人宫颈癌 细胞HeLa 的研究.

Objective To observe the proliferation, metastasis, invasion, angiogenesis and the changes of PI3K/AKT signaling pathway after transfection of human cervical cancer cell HeLa with pcDNA-STC1 carried by ultrasound microbubble contrast agent. Methods Using GEPIA database to analyze the expression level of STC1 in cervical cancer tissue, the patient's Prognosis and survival time. 80 cases of cervical cancer patients diagnosed and treated in our hospital from January to October 2019 were selected as research objects, cancer tissues and adjacent tissues were collected, and the expression of STC1 mRNA was detected by qRT-PCR. The relationship between pathological factors; HeLa cell grouping: Blank control group: HeLa cells were cultured normally without any treatment. Negative control group (HeLa-pcDNA-NC): 5% microvesicle concentration + 2 μg empty plasmid + serum-free DMEM medium .Experimental group (HeLa-pcDNA-STC1): 5% microbubble concentration + 2 μg pcDNA-STC1 plasmid + serum-free DMEM medium. MTT method and the number of single colonies formed by cancer cells were used to detect cell proliferation; scratch test to detect cell migration ;Transwell chamber test to detect cell invasion ability; Matrigel in vitro tube formation test to detect cell angiogenesis ability; Western blot to detect p-PI3K, p-AKT, matrix metalloproteinase-2 (MMP-2)/matrix metalloproteinase-9 ( MMP-9), vascular endothelial growth factor (VEGF), and angiopoietin 2 (Ang2). Results GEPIA database showed that the expression level of STC1 in cervical cancer tissue was lower than that in normal tissue, and the prognosis and survival time were negatively correlated with the expression level of STC1. Correlation; compared with adjacent normal cervical tissue, cervical cancer tissue and STC1 mRNA were down-regulated, and miR-574-5p level was negatively correlated with STC1 level, which was significantly correlated with tumor differentiation, FIGO stage, and lymph node metastasis. Compared with the blank control group and the negative control group, the OD value of the experimental group decreased at 48 h, 72 h, and 96 h, the number of cell clones was extremely small, the migration rate decreased, the number of cell invasion decreased, and the angiogenesis ability decreased, and MMP-2, MMP- 9. The expression of VEGF, Ang2, p-PI3K, and p-AKT proteins was down-regulated (P<0.05). Conclusion Ultrasonic microbubble contrast agent carrying pcDNA-STC1 transfection can significantly inhibit the proliferation, metastasis, invasion and angiogenesis of human cervical cancer cell HeLa, and its mechanism of action may be achieved by inhibiting the PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]