舒芬太尼调控 miR-1抑制肝癌细胞 MHCC97-H增殖、侵袭和迁移的机制研究

目的 探讨舒芬太尼调控微小RNA-1(miR-1)表达对人肝癌MHCC97-H细胞增殖、侵袭和迁移的影响及其机制.方法 本研究起止时间为2018年3月至2019年9月,采用细胞计数试剂盒(CCK-8)实验检测不同浓度的舒芬太尼对人肝癌MHCC97-H细胞活力的影响,qRT-PCR检测舒芬太尼对MHCC97-H细胞中miR-1表达的影响;通过脂质体转染法将miR-1抑制剂(inhibitor)及阴性对照转染至MHCC97-H细胞,实验分为Control组、舒芬太尼(Sufentanil)组、转染对照(Sufentanil+NC)组和转染(Sufent-anil+miR-1)组.细胞计数试剂盒(CCK-8)实验检测各组MHCC97-H细胞增殖能力,Transwell实验检测各组MHCC97-H细胞侵袭能力,划痕实验检测各组MHCC97-H细胞迁移能力,蛋白质印迹法(Western blotting)检测各组MHCC97-H细胞中基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)蛋白表达情况.结果 不同浓度的舒芬太尼对肝癌MHCC97-H细胞活力具有不同程度的抑制作用;与Control组比,Sufentanil组MHCC97-H细胞中miR-1的表达(1.00±0.09)比(2.25±0.24)]明显升高(P<0.05),细胞增殖(0.54±0.05)比(0.34±0.03)]、侵袭(106.24±8.25)个比(51.34±5.02)个]和迁移能力(37.36±4.01)％比(16.34±1.72)％]降低(P<0.05),MMP-2(0.33±0.04)比(0.15±0.02)]和MMP-9(0.24±0.02)比(0.11±0.01)]的表达下调(P<0.05);与Sufentanil+NC组比,Sufentanil+miR-1组MHCC97-H细胞中miR-1的表达(2.20±0.21)比(1.42±0.15)]明显降低(P<0.05),细胞增殖(0.32±0.03)比(0.46±0.04)]、侵袭(52.36±5.14)个比(91.16±6.06)个]和迁移能力(17.11±1.71)％比(29.85±3.10)％]升高(P<0.05),MMP-2(0.15±0.02)比(0.27±0.02)]和MMP-9(0.13±0.01)比(0.19±0.02)]的表达上调(P<0.05).结论 舒芬太尼可通过调控miR-1的表达抑制人肝癌MHCC97-H细胞的增殖、侵袭和迁移,其机制可能与调控MMP-2和MMP-9表达有关.

Mechanism of sufentanil inhibiting proliferation, invasion and migration of hepatocellular carcinoma cell line MHCC97-H by regulating miR-1

Objective To explore the effect and mechanism of sufentanil on the proliferation, invasion and migration of human hepatocellular carcinoma cell line MHCC97-H by regulating microRNA-1 (miR-1) expression.Methods The start and end time of this research was from March 2018 to September 2019.The effects of different concentrations of sufentanil on the viability of human hepatocellular carcinoma cell line MHCC97-H were determined using the cell counting kit-8 (CCK-8) assay. The effect of sufentanil on the expression of miR-1 in MHCC97-H cells was detected by qRT-PCR. The miR-1 inhibitor and negative control were transfected into MHCC97-H cells using liposome-mediated transfection. There were control group, sufentanil group, sufentanil+NC group and sufentanil+miR-1 group in this study. The proliferation, invasion and migration ability of MHCC97-H cells in each group was detected by CCK-8 assay, Transwell assay, and scratch test, respectively. Western blotting was used to detect the expressions of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in MHCC97-H cells.Results Different concentrations of sufentanil had different inhibitory effects on the viability of MHCC97-H cells in vitro. Compared with the control group, the expression of miR-1 in MHCC97-H cells in the sufentanil group (1.00±0.09) vs. (2.25±0.24)] was significantly higher (P<0.05), the cell proliferation (0.54± 0.05) vs. (0.34±0.03)], invasion (106.24±8.25) vs. (51.34±5.02)] and migration ability (37.36±4.01)% vs. (16.34±1.72)%] decreased (P <0.05), and MMP-2 (0.33±0.04) vs. (0.15±0.02)] and MMP-9 (0.24±0.02) vs. (0.11±0.01)] expressions were down-regulated (P<0.05). Compared with sufentanil+NC group, the expression of miR-1 in MHCC97-H cells in the sufentanil+miR-1 group (2.20±0.21) vs. (1.42±0.15)] was significantly reduced (P<0.05), cell proliferation (0.32±0.03) vs. (0.46±0.04)], invasion (52.36±5.14) vs. (91.16± 6.06)] and migration ability (17.11±1.71)% vs. (29.85±3.10)%] increased (P<0.05), and MMP-2 (0.15±0.02) vs. (0.27±0.02)] and MMP-9 (0.13±0.01) vs. (0.19±0.02)] expressions were up-regulated (P<0.05).Conclusion Sufentanil can inhibit the proliferation, invasion and migration of human hepatocellular carcinoma cell line MHCC97-H by regulating the expression of miR-1, whose mechanism may be related to the regulation of MMP-2 and MMP-9 expressions.