中国临床药理学杂志

咪达唑仑调节LKB1/AMPK信号通路对结直肠癌细胞增殖、侵袭及凋亡影响的研究

PDF下载

徐国兴 ^1a , 徐国亭 ^1a , 王苗级 ^1a , 刘耿 ^1b

中国临床药理学杂志 | 临床与基础桥接研究 2025,41(21): 3103-3109

收起

中国临床药理学杂志 |临床与基础桥接研究 2025 , 41 (21) : 3103 -3109

咪达唑仑调节LKB1/AMPK信号通路对结直肠癌细胞增殖、侵袭及凋亡影响的研究

全屏

徐国兴^1a, 徐国亭^1a, 王苗级^1a, 刘耿^1b

作者信息

^1a.南阳医学高等专科学校第一附属医院，麻醉与围术期医学科，河南南阳 473000

^1b.南阳医学高等专科学校第一附属医院，普通外科，河南南阳 473000

通讯作者:

徐国兴 Tel: （0377）63328360 E-mail: bvxzzn@163.com

作者简介:

徐国兴（1984-），男，副主任医师，从事临床麻醉方面的研究和工作

Research of midazolam on the proliferation, invasion and apoptosis of colorectal cancer cells by regulating the LKB1/AMPK signaling pathway

Guo-xing XU^1a, Guo-ting XU^1a, Miao-ji WANG^1a, Geng LIU^1b

Affiliations

^1a.Department of Anesthesiology and Perioperative Medicine, the First Affiliated Hospital of Nanyang Medical College, Nanyang 473000, Henan Province, China

^1b.Department of General Surgery, the First Affiliated Hospital of Nanyang Medical College, Nanyang 473000, Henan Province, China

出版时间: 2025-11-17 doi: 10.13699/j.cnki.1001-6821.2025.21.017

文章导航

摘要

收起

目的

探讨咪达唑仑调节肝激酶B1（LKB1）/腺苷酸（AMP）活化蛋白激酶（AMPK）信号通路对结直肠癌细胞增殖、侵袭及凋亡的影响。

方法

用细胞计数试剂盒-8（CCK-8）实验法检测不同浓度（5、10、20和40 μg·mL^-1）的咪达唑仑对HCT116细胞增殖的影响，选择20 μg·mL^-1咪达唑仑进行后续实验。将HCT116细胞分为空白组（不作处理）、咪达唑仑组（20 μg·mL^-1咪达唑仑）、si-NC组（转染si-NC质粒）、si-LKB1组（转染si-LKB1质粒）、咪达唑仑+ si-NC组（转染si-NC质粒+20 μg·mL^-1咪达唑仑）和咪达唑仑+ si-LKB1组（转染si-LKB1质粒+20 μg·mL^-1咪达唑仑）。用流式细胞法、细胞计数试剂盒-8（CCK-8）实验法、EdU试剂盒法及Transwell实验法检测细胞凋亡、增殖及侵袭变化；用实时荧光定量逆转录聚合酶链反应（qRT-PCR）检测LKB1和AMPK mRNA相对表达水平；用蛋白质印迹法检测增殖细胞核抗原（PCNA）、半胱氨酸天冬氨酸蛋白酶3（Caspase-3）、金属蛋白酶9（MMP-9）、LKB1和AMPK蛋白相对表达水平。

结果

空白组、咪达唑仑组、si-NC组、si-LKB1组、咪达唑仑+si-NC组和咪达唑仑+si-LKB1组的EdU阳性率分别为（62.52±6.38）%、（35.26±3.64）%、（61.95±6.24）%、（88.63±8.94）%、（36.05±3.71）%和（55.62±5.67）%，增殖率分别为（90.85±9.04）%、（48.85±4.92）%、（91.04±8.92）%、（129.32±13.05）%、（49.05±4.98）%和（79.86±8.11）%，凋亡率分别为（5.05±0.53）%、（20.22±2.15）%、（4.86±0.49）%、（3.11±0.35）%、（19.86±2.08）%和（8.77±0.91）%，侵袭数分别为198.64±20.45、122.53±12.37、195.32±20.34、286.61±29.33、123.64±12.46和176.64±18.06，LKB1 mRNA相对表达水平分别为0.93±0.10、2.86±0.31、1.08±0.12、0.57±0.07、2.78±0.29和1.55±0.17，AMPK mRNA相对表达水平分别为1.06±0.11、2.25±0.24、0.98±0.10、0.42±0.05、2.19±0.23和1.35±0.15，PCNA相对表达水平分别为0.79±0.08、0.42±0.05、0.81±0.09、1.22±0.14、0.46±0.05和0.68±0.07，Caspase-3相对表达水平分别为0.92±0.10、1.44±0.16、1.03±0.11、0.51±0.06、1.48±0.16和1.07±0.11，MMP-9相对表达水平分别为0.98±0.10、0.52±0.06、1.01±0.12、1.53±0.17、0.58±0.06和0.82±0.09，LKB1相对表达水平分别为0.31±0.03、0.76±0.08、0.34±0.04、0.15±0.02、0.72±0.08和0.45±0.05，AMPK相对表达水平分别为0.44±0.05、0.85±0.09、0.42±0.05、0.23±0.03、0.80±0.09和0.57±0.06，咪达唑仑组的上述指标与空白组比较；si-LKB1组的上述指标与si-NC组比较；咪达唑仑+si-LKB1组的上述指标与咪达唑仑+si-NC组及si-LKB1组比较，在统计学上差异均有统计学意义（均P<0.05）。

结论

咪达唑仑激活LKB1/AMPK信号通路抑制结直肠癌细胞增殖、侵袭，诱导其凋亡。

关键词

咪达唑仑 / 结直肠癌 / 肝激酶/腺苷酸活化蛋白激酶信号通路 / 侵袭 / 增殖 / 凋亡

Abstract

收起

Objective

To explore the effects of midazolam on the proliferation, invasion and apoptosis of colorectal cancer cells by regulating the liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK) signaling pathway.

Methods

Cell counting kit-8 method was used to detect the effects of different concentrations (5, 10, 20 and 40 μg·mL^-1) midazolam on the proliferation of HCT116 cells. 20 μg·mL^-1 midazolam was selected for the subsequent experiments. HCT116 cells were grouped into blank group (no treatment), midazolam group (20 μg·mL^-1 midazolam), si-NC group (transfected with si-NC plasmid), si-LKB1 group (transfected with si-LKB1 plasmid), midazolam+si-NC group (transfected with si-NC plasmid+20 μg·mL^-1 midazolam) and midazolam+si-LKB1 group (transfected with si-LKB1 plasmid+20 μg·mL^-1 midazolam). Flow cytometry, cell countig kit-8 (CCK-8), EdU and Transwell experiments were used to detect changes in cell apoptosis, proliferation and invasion. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure the LKB1 and AMPK mRNAs. Western blot was used to measure the proliferating cell nuclear antigen (PCNA), cysteinyl aspartate-specific protease 3 (caspase-3), matrix metalloproteinase 9 (MMP-9), liver kinase B1 (LKB1) and AMP-activated protein kinase (AMPK) proteins.

Results

The EdU positive rates of the blank group, midazolam group, si-NC group, si-LKB1 group, midazolam + si-NC group and midazolam + si-LKB1 group were (62.52±6.38)%, (35.26±3.64)%, (61.95±6.24)%, (88.63±8.94)%, (36.05±3.71)% and (55.62±5.67)%, respectively; the proliferation rates were (90.85±9.04)%, (48.85±4.92)%, (91.04±8.92)%, (129.32±13.05)%, (49.05±4.98)% and (79.86±8.11)%, respectively; the apoptosis rates were (5.05±0.53)%, (20.22±2.15)%, (4.86±0.49)%, (3.11±0.35)%, (19.86±2.08)% and (8.77±0.91)%, respectively; the invasion numbers were 198.64±20.45, 122.53±12.37, 195.32±20.34, 286.61±29.33, 123.64±12.46 and 176.64±18.06, respectively; the relative expression levels of LKB1 mRNA were 0.93±0.10, 2.86±0.31, 1.08±0.12, 0.57±0.07, 2.78±0.29 and 1.55±0.17, respectively; AMPK mRNA were 1.06±0.11, 2.25±0.24, 0.98±0.10, 0.42±0.05, 2.19±0.23 and 1.35±0.15, respectively; the relative protein expression levels of PCNA were 0.79±0.08, 0.42±0.05, 0.81±0.09, 1.22±0.14, 0.46±0.05 and 0.68±0.07, respectively; Caspase-3 were 0.92±0.10, 1.44±0.16, 1.03±0.11, 0.51±0.06, 1.48±0.16 and 1.07±0.11, respectively; MMP-9 were 0.98±0.10, 0.52±0.06, 1.01±0.12, 1.53±0.17, 0.58±0.06 and 0.82±0.09, respectively; LKB1 were 0.31±0.03, 0.76±0.08, 0.34±0.04, 0.15±0.02, 0.72±0.08 and 0.45±0.05, respectively; AMPK were 0.44±0.05, 0.85±0.09, 0.42±0.05, 0.23±0.03, 0.80±0.09 and 0.57±0.06, respectively. For all these indicators, compared between the midazolam group and the blank group, the si-LKB1 group and the si-NC group, the midazolam + si-LKB1 group and the midazolam + si-NC group, and the si-LKB1 group, all showed statistically significant differences (all P<0.05).

Conclusion

Midazolam activates the LKB1/AMPK signaling pathway, inhibits the proliferation and invasion of colorectal cancer cells, and induces their apoptosis.

Key words

midazolam / colorectal cancer / liver kinase B1/ adenosine monophosphate activated protein kinase signaling pathway / invasion / proliferation / apoptosis

引用本文

徐国兴, 徐国亭, 王苗级, 刘耿. 咪达唑仑调节LKB1/AMPK信号通路对结直肠癌细胞增殖、侵袭及凋亡影响的研究. 中国临床药理学杂志, 2025 , 41 (21) : 3103 -3109 . DOI: 10.13699/j.cnki.1001-6821.2025.21.017

Guo-xing XU, Guo-ting XU, Miao-ji WANG, Geng LIU. Research of midazolam on the proliferation, invasion and apoptosis of colorectal cancer cells by regulating the LKB1/AMPK signaling pathway[J]. Chinese Journal of Clinical Pharmacology, 2025 , 41 (21) : 3103 -3109 . DOI: 10.13699/j.cnki.1001-6821.2025.21.017

基金

收起

河南省医学科技攻关计划项目基金资助项目(LHGJ20210104)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

2025年第41卷第21期

PDF下载

132

引用本文

BibTeX

文章信息

doi: 10.13699/j.cnki.1001-6821.2025.21.017

接收时间：2025-07-16
首发时间：2026-08-05
出版时间：2025-11-17

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2025-07-16

基金

河南省医学科技攻关计划项目基金资助项目(LHGJ20210104)

作者信息

^1a.南阳医学高等专科学校第一附属医院，麻醉与围术期医学科，河南南阳 473000

^1b.南阳医学高等专科学校第一附属医院，普通外科，河南南阳 473000

通讯作者:

徐国兴 Tel: （0377）63328360 E-mail: bvxzzn@163.com

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zglcylxzz/CN/10.13699/j.cnki.1001-6821.2025.21.017

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT