超声介导载miR-16-5p微泡联合树突状细胞疫苗对小鼠肝癌皮下移植瘤生长的影响*

doi:10.3969/j.issn.1672-5069.2022.01.005

摘要/Abstract

摘要： 目的研究采用超声介导载miR-16-5p微泡联合树突状细胞(DC)疫苗处理对小鼠肝癌皮下移植瘤生长的影响。方法制备载miR-16-5p的高分子微泡和DC疫苗。在30只小鼠用HepG2细胞建立肝癌皮下移植瘤模型,再随机分为对照组、空微泡、miR-16-5p微泡、DC疫苗和miR-16-5p微泡联合DC疫苗处理组,每组6只。观察各组小鼠肿瘤生长情况,绘制肿瘤生长曲线并计算抑瘤率,采用实时荧光定量RT-PCR法检测肿瘤组织miR-16-5p水平。采用ELISA法检测血清免疫球蛋白G(IgG)、IFN-γ、TGF-β、IL-2和IL-10水平。结果制备的高分子微泡呈球形,平均粒径为1.31 μm;对照组、空微泡、miR-16-5p微泡、DC疫苗和联合处理组小鼠肿瘤体积分别为(5973.2±257.4)mm³、(5912.3±248.3)mm³、(4873.7±281.5)mm³、(4954.9±276.3)mm³和(2225.8±201.4)mm³,瘤体质量分别为(4.4±0.4)g、(4.2±0.3)g、(2.8±0.2)g、(2.7±0.2)g和(0.9±0.2)g,差异显著(P<0.05);各组小鼠肿瘤组织miR-16-5p水平分别为(1.02±0.06)、(1.05± 0.03)、(7.81±0.97)、(1.08±0.07)和(8.08± 1.06),提示微泡组和联合组显著高于其他各组(P<0.05);与对照组或空微泡处理组比,其他三组血清IgG、IFN-γ和IL-2水平均显著升高,其中联合组水平最高,而其他三组血清TGF-β和IL-10水平均显著降低,其中联合组水平最低(均P<0.05)。结论采用超声介导载miR-16-5p微泡联合DC疫苗处理对小鼠肝癌皮下移植瘤生长具有协同的抑制作用,可能与增强了小鼠对肿瘤的免疫效应有关。

关键词: 肝癌皮下移植瘤, 超声介导载微泡, 微小RNA-16-5p, 树突状细胞疫苗, 小鼠

Abstract: Objective The paper aimed to investigate the effect of ultrasound-mediated miR-16-5p loaded microbubbles and dendritic cell (DC) vaccine on subcutaneous transplanted tumor growth in mice. Methods The polymer microbubbles loaded with miR-16-5p were prepared and their general properties were detected. The subcutaneous transplanted tumors by HepG2 cell injection were established in thirty mice, and were randomly divided into control, vacuous microbubble, miR-16-5p microbubble, DC vaccine, and miR-16-5p microbubble and DC vaccine combination intervention groups, with six mice in each group. The tumor growth of mice in each group was observed, the tumor growth curve was drawn and the tumor inhibition rate was calculated. The miR-16-5p levels in tumorous tissues were detected by real-time RT-PCR. Serum levels of IgG, IFN-γ, TGF-β, IL-2, and IL-10 were measured by ELISA. Results The prepared polymer microbubbles were spherical with an average particle size of 1.31 μm; the tumor volumes in control, vacuous microbubble, miR-16-5p microbubble, DC vaccine, and miR-16-5p microbubble and DC vaccine combination-intervened groups were(5973.2±257.4)mm³, (5912.3±248.3)mm³, (4873.7±281.5)mm³, (4954.9±276.3)mm³ and (2225.8±201.4)mm³, and the tumor mass were (4.4±0.4)g, (4.2±0.3)g, (2.8±0.2)g, (2.7±0.2)g and (0.9±0.2)g, significantly different among them(P<0.05); the tumorous miR-16-5p levels were (1.02±0.06), (1.05± 0.03), (7.81±0.97), (1.08±0.07) and (8.08± 1.06), suggesting that those in miR-16-5p microbubble- and DC vaccine combination-intervened groups were significantly higher than in other three groups (P<0.05); serum IgG, IFN-γ and IL-2 levels significantly increased, while serum TGF-β and IL-10 levels greatly decrease in miR-16-5p microbubble, DC vaccine, and miR-16-5p microbubble and DC vaccine combination intervention groups as compared to those in control or vacuous microbubble-intervened groups (P<0.05). Conclusion The administration of ultrasound-mediated miR-16-5p microbubbles and DC vaccine has a synergistic inhibitory effect on the growth of subcutaneous transplanted hepatoma in mice, which might be related to the enhancement immune functions.

Key words: Transplanted tumor, Ultrasound-mediated loaded microbubbles, miR-16-5p, Dendritic cell vaccine, Mice

时牛, 宋洁, 崔天庆, 云宇婷, 郭瑞芳, 吴昆. 超声介导载miR-16-5p微泡联合树突状细胞疫苗对小鼠肝癌皮下移植瘤生长的影响^*[J]. 实用肝脏病杂志, 2022, 25(1): 17-21.

Shi Niu, Song Jie, Cui Tianqing, et al. Inhibition of subcutaneous transplanted hepatoma growth by ultrasound-mediated miR-16-5p loaded microbubbles with dendritic cell vaccine combination in mice[J]. Journal of Practical Hepatology, 2022, 25(1): 17-21.

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超声介导载miR-16-5p微泡联合树突状细胞疫苗对小鼠肝癌皮下移植瘤生长的影响^*

Inhibition of subcutaneous transplanted hepatoma growth by ultrasound-mediated miR-16-5p loaded microbubbles with dendritic cell vaccine combination in mice

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