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香叶木素调控STX17-AS1/miR-383-5p轴对结直肠癌细胞增殖、凋亡和迁移的影响
Effect of diosmetin on proliferation, apoptosis and migration of colorectal cancer cells by regulating STX17-AS1/miR-383-5p axis
- 第三军医大学学报 2021年43卷第17期 页码：1642-1649
- 作者机构：
- 作者简介：
- 基金信息：
DOI：10.16016/j.1000-5404.202103147
中图分类号：收稿：2021-03-18，
纸质出版：2021-09-15
- 稿件说明：
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吴远, 刘吉宇, 张琴. 香叶木素调控STX17-AS1/miR-383-5p轴对结直肠癌细胞增殖、凋亡和迁移的影响[J]. 第三军医大学学报, 2021,43(17):1642-1649.
Yuan WU, Jiyu LIU, Qin ZHANG. Effect of diosmetin on proliferation, apoptosis and migration of colorectal cancer cells by regulating STX17-AS1/miR-383-5p axis[J]. Journal of Third Military Medical University, 2021, 43(17): 1642-1649.
吴远, 刘吉宇, 张琴. 香叶木素调控STX17-AS1/miR-383-5p轴对结直肠癌细胞增殖、凋亡和迁移的影响[J]. 第三军医大学学报, 2021,43(17):1642-1649. DOI： 10.16016/j.1000-5404.202103147.
Yuan WU, Jiyu LIU, Qin ZHANG. Effect of diosmetin on proliferation, apoptosis and migration of colorectal cancer cells by regulating STX17-AS1/miR-383-5p axis[J]. Journal of Third Military Medical University, 2021, 43(17): 1642-1649. DOI： 10.16016/j.1000-5404.202103147.
摘要
目的
2
探讨香叶木素对结直肠癌细胞增殖、凋亡和迁移的影响，分析其机制是否与调控STX17-AS1/miR-383-5p轴有关。
方法
2
通过细胞计数试剂盒（CCK-8）、集落形成实验、划痕愈合实验、流式细胞术、实时定量PCR（RT-qPCR）分析香叶木素（10、20、40 μmol/L）对结直肠癌细胞HCT116的活力、集落形成数、划痕愈合率、凋亡以及STX17-AS1、miR-383-5p表达的影响。RT-qPCR分析2019年本院29例结直肠癌患者癌组织和对应癌旁组织中STX17-AS1、miR-383-5p相对水平。双荧光素酶报告实验分析STX17-AS1和miR-383-5p的靶向关系。分别转染STX17-AS1的小干扰RNA、miR-383-5p模拟物至HCT116细胞，观察抑制STX17-AS1或过表达miR-383-5p对HCT116细胞增殖、迁移和侵袭的影响。
结果
2
香叶木素（10、20、40 μmol/L）处理显著降低HCT116细胞活力、集落形成数、划痕愈合率（
P
<
0.05），增加细胞凋亡率（
P
<
0.05），降低STX17-AS1表达水平（
P
<
0.05），增加miR-383-5p表达水平（
P
<
0.05）。结直肠癌组织中STX17-AS1的表达水平较癌旁组织显著升高（
P
<
0.05），miR-383-5p的表达水平较癌旁组织显著降低（
P
<
0.05）。STX17-AS1与miR-383-5p直接结合。抑制STX17-AS1表达显著降低HCT116细胞活力、集落形成数、划痕愈合率（
P
<
0.05），增加细胞凋亡率（
P
<
0.05），增加miR-383-5p表达水平（
P
<
0.05）。过表达miR-383-5p显著降低HCT116细胞活力、集落形成数、划痕愈合率（
P
<
0.05），增加细胞凋亡率（
P
<
0.05）。过表达STX17-AS1或抑制miR-383-5p表达显著减弱香叶木素对HCT116细胞活力、集落形成数、划痕愈合率和凋亡率的影响（
P
<
0.05）。
结论
2
香叶木素通过下调STX17-AS1/miR-383-5p轴可抑制结直肠癌细胞增殖和迁移，诱导细胞凋亡。
Abstract
Objective
2
To investigate the effect of diosmetin on the proliferation
apoptosis and migration of colorectal cancer cells
and further to analyze whether its mechanism is related to the regulation of STX17-AS1/miR-383-5p axis.
Methods
2
Cell counting kit (CCK-8) assay
colony formation test
scratch healing test
flow cytometry and real-time quantitative PCR (RT-qPCR) were used to analyze the effects of diosmetin (10
20 and 40 μmol/L) on cell viability
colony formation
scratch healing rate
apoptosis
and STX17-AS1 and miR-383-5p expression in HCT116 cells. The relative expression levels of STX17-AS1 and miR-383-5p in cancer tissues and adjacent tissues of 29 patients with colorectal cancer in our hospital were assessed using RT-qPCR. Dual luciferase reporter assay system was employed to analyze the targeting relationship between STX17-AS1 and miR-383-5p. The small interfering RNA of STX17-AS1 and miR-383-5p mimics of STX17-AS1 were transfected into HCT116 cells respectively
and the effects of STX17-AS1 inhibition or miR-383-5p overexpression on the proliferation
apoptosis and migration of HCT116 cells were also analyzed.
Results
2
Diosmetin treatment (10
20
40 μmol/L) significantly reduced cell viability
colony formation numbers and scratch healing rate (
P
<
0.05)
increased cell apoptotic rate (
P
<
0.05)
decreased STX17-AS1 expression level (
P
<
0.05)
and increased the expression level of miR-383-5p in HCT116 cells (
P
<
0.05). The expression level of STX17-AS1 was significantly higher (
P
<
0.05)
and that of miR-383-5p was significantly lower in the colorectal cancer tissues than the adjacent tissues (
P
<
0.05). STX17-AS1 directly bound to miR-383-5p. Inhibition of STX17-AS1 significantly reduced cell viability
colony formation numbers and scratch healing rate (
P
<
0.05)
increased cell apoptotic rate (
P
<
0.05)
and enhanced the miR-383-5p expression level in HCT116 cells (
P
<
0.05). While
miR-383-5p overexpression si
gnificantly reduced cell viability
colony formation numbers and scratch healing rate (
P
<
0.05)
and increased cell apoptosis rate in the cells (
P
<
0.05). Overexpression of STX17-AS1 or inhibition of miR-383-5p significantly weakened the effect of diosmetin on the above indicators of HCT116 cells (
P
<
0.05).
Conclusion
2
Diosmetin inhibits the proliferation and migration of colorectal cancer cells and induces apoptosis by down-regulating the STX17-AS1/miR-383-5p axis
关键词
Keywords
references
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