miRNA Expression of SMMC-7721 Cell Induced by Xiaoai Jiedu Formula

XU Li-li; CHEN Hui; WU Ming-jie; CHEN Hai-bin; LI Wen-ting; SHEN Zheng-jie; SHANGGUAN Duan-dan; DONG Fan; LI Lin-wan; ZHOU Hong-guang

doi:10.13422/j.cnki.syfjx.20180616

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miRNA Expression of SMMC-7721 Cell Induced by Xiaoai Jiedu Formula

更新时间：2024-01-04

- miRNA Expression of SMMC-7721 Cell Induced by Xiaoai Jiedu Formula
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 7, Pages: 89-94(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20180616
  CLC： R285
- Published：2018
- 稿件说明：
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XU Li-li, CHEN Hui, WU Ming-jie, et al. miRNA Expression of SMMC-7721 Cell Induced by Xiaoai Jiedu Formula[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(7): 89-94.
DOI：

XU Li-li, CHEN Hui, WU Ming-jie, et al. miRNA Expression of SMMC-7721 Cell Induced by Xiaoai Jiedu Formula[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(7): 89-94. DOI： 10.13422/j.cnki.syfjx.20180616.

摘要

目的：探讨消癌解毒方对人肝癌SMMC-7721细胞增殖及微小核糖核酸（microRNA，miR）-25-3p，miR-29a-5p，miR-122-3p，miR-124-3p，miR-182-5p表达谱的影响。方法：将12只雄性大耳白兔随机分为4组，分别为消癌解毒方高、中、低剂量（15.12，7.56，3.78 g ·kg-1）组和空白组，各组分别给予消癌解毒方及生理盐水灌胃4 d。4 d后于颈动脉中取血清并配制培养基。用消癌解毒方（15.12，7.56，3.78 g ·kg-1）含药血清及空白血清的培养基处理人肝癌细胞株SMMC-7721，噻唑蓝（MTT）比色法检测肿瘤细胞增殖活性、实时荧光定量逆转录聚合酶链式反应（Real-time polymerase chain reaction，Real-time PCR）法验证人肝癌细胞SMMC-7721中miR-25-3p，miR-29a-5p，miR-122-3p，miR-124-3p和miR-182-5p的表达水平。结果：经过12，24，48 h后，消癌解毒方（15.12，7.56，3.78 g ·kg-1）含药血清对人肝癌SMMC-7721细胞的增殖均存在抑制作用。随着消癌解毒方含药血清浓度的增加，其对人肝癌细胞SMMC-7721细胞增殖的抑制率也相应增加。其中高剂量组含药血清的抑制效果最好，其干预12，48 h的吸光度A较同期空白组明显降低（P<0.05）。高剂量组干预24 h A比同期空白组显著降低（P<0.01），该组抑制率为42.86%。Real-time PCR证实消癌解毒方（15.12，7.56，3.78 g ·kg-1）含药血清的培养基能诱导miR-25-3p，miR-182-5p表达下调，诱导miR-29a-5p，miR-122-3p，miR-124-3p表达上调。且高剂量组的调控作用较空白组更显著（P<0.01）。结论：消癌解毒方可能通过诱导miRNA表达谱的改变而参与抑制人肝癌细胞SMMC-7721增殖作用，但其具体机制仍有待进一步研究。

Abstract

Objective: To study the effect of Xiaoai Jiedu formula on the proliferation of human hepatocarcinoma SMMC-7721 cells and expression profiles of microRNA(miR)-25-3p

miR-29a-5p

miR-122-3p

miR-124-3p and miR-182-5p. Method: Twelve male white rabbits were randomly divided into four groups. They were respectively given high-dose

middle-dose and low-dose Xiaoai Jiedu formula or normal saline for four days. Serum was taken from their carotid blood and then made into culture medium. Cytotoxicity of Xiaoai Jiedu formula against SMMC-7721 cells was measured by 3-(4

5-dimethyl-2-thiazolyl)-2

5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Expression profiles of miR-25-3p

miR-29a-5p

miR-122-3p

miR-124-3p and miR-182-5p of SMMC-7721 cells were analyzed by a miRNA array and Real-time polymerase chain reaction (Real-time PCR). Result: High-dose

middle-dose and low-dose Xiaoai Jiedu formula could all inhibit the proliferation of SMMC-7721 cells after 12

48 h. Xiaoai Jiedu formula gradually increased the proliferation inhibition rate of SMMC-7721 cells in a dose-dependent manner

with the increase of drug concentration. The absorbancy of the high-dose group showed statistical differences after 12 h and 48 h compared with blank control group at the same time points (P<0.05). The absorbancy of the high-dose group showed statistically significant differences after 24 h compared with the blank control group at the same time point (P<0.01). The absorbancy of the high dose group after intervention for 24 h was significantly lower than that of blank control group (P<0.01)

the group's inhibition rate was 42.86%. Moreover

Real-time PCR verified the down-regulation of miR-25-3p

miR-182-5p and the up-regulation of miR-29a-5p

miR-122-3p

miR-124-3p induced by Xiaoai Jiedu formula. The regulatory effect of the high-dose group was more significant than that of the blank control group (P<0.01). Conclusion: Xiaoai Jiedu formula may inhibit the proliferation of SMMC-721 by inducing the change in microRNA (miRNA) expression. However

the concrete mechanism remains to be further studied.

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