[1]柏希慧,刘诗雨,孙媛媛.拉罗替尼通过AMPK/mTOR信号通路调控结肠癌细胞自噬和抑制增殖与迁移的实验研究[J].现代检验医学杂志,2024,39(06):29-36.[doi:10.3969/j.issn.1671-7414.2024.06.005]
　BAI Xihui,LIU Shiyu,SUN Yuanyuan.Experimental Study of Larotrectinib Regulating Autophagy and Inhibiting Proliferation and Migration of Colon Cancer Cells Through AMPK/mTOR Signaling Pathway[J].Journal of Modern Laboratory Medicine,2024,39(06):29-36.[doi:10.3969/j.issn.1671-7414.2024.06.005]
点击复制

拉罗替尼通过AMPK/mTOR信号通路调控结肠癌细胞自噬和抑制增殖与迁移的实验研究()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年06期
页码:
29-36
栏目:
论著
出版日期:
2024-11-15

文章信息/Info

Title:
Experimental Study of Larotrectinib Regulating Autophagy and Inhibiting Proliferation and Migration of Colon Cancer Cells Through AMPK/mTOR Signaling Pathway
文章编号:
1671-7414（2024）06-029-08
作者:
柏希慧1; 刘诗雨2; 孙媛媛1
（1. 西安交通大学第一附属医院药学部，西安 710061；2. 西安市阎良区人民医院，西安 710089）
Author(s):
BAI Xihui1; LIU Shiyu2; SUN Yuanyuan1
（1.Department of Pharmacy, the First Affiliated Hospital of Xi’an Jiaotong University，Xi’an 710061，China；2. People’s Hospital of Yanliang District，Xi’an 710089，China）
关键词:
结肠癌; 拉罗替尼; 腺苷酸活化蛋白激酶/ 哺乳动物雷帕霉素靶蛋白通路; 自噬
分类号:
R735.35；R730.43
DOI:
10.3969/j.issn.1671-7414.2024.06.005
文献标志码:
A
摘要:
（1. 西安交通大学第一附属医院药学部，西安 710061；2. 西安市阎良区人民医院，西安 710089）
Abstract:
Objective To investigate the effects of Larotrectinib (Lar) on autophagy, proliferation and migration of colon cancer cells and its molecular mechanism. Methods Human colon cancer cell lines COLO 205, HCT 116 and human colonic epithelial cell line CP-H040 were treated with different concentrations of Lar (0, 100, 200, 400, 800, 1600 and 3 200 nmol/L). CCK-8 assay was used to detect the cell viability of COLO 205, HCT 116 and CP-H040 cells. COLO 205 and HCT 116 cells were randomly divided into control group (Con group), Lar group, Chloroquine group (CQ group) and Lar+CQ group. Cell invasion was detected by Transwell assay. Scratch test was used to detect cell migration ability. Ki67 immunofluorescence staining was used to detect cell proliferation. Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of epithelialmesenchymal transition related markers in colon cancer cells. Autophagy was detected by adenovirus transfection experiment and transmission electron microscopy. Western blot was used to detect the expression of adenosine monophosphate-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) pathway related proteins. Results Lar significantly inhibited the viability of COLO 205 and HCT 116 cells in a concentration-dependent manner (F=355.181, 403.758, all P<0.001). Compared with the Con group, the number of invasive cells, Ki67 fluorescence intensity and scratch healing rate of colon cancer cells in the Lar group were decreased, the expression of E-cadherin mRNA was increased, the expressions of Vimentin and MMP2 mRNA Objective To investigate the effects of Larotrectinib (Lar) on autophagy, proliferation and migration of colon cancer cells and its molecular mechanism. Methods Human colon cancer cell lines COLO 205, HCT 116 and human colonic epithelial cell line CP-H040 were treated with different concentrations of Lar (0, 100, 200, 400, 800, 1600 and 3 200 nmol/L). CCK-8 assay was used to detect the cell viability of COLO 205, HCT 116 and CP-H040 cells. COLO 205 and HCT 116 cells were randomly divided into control group (Con group), Lar group, Chloroquine group (CQ group) and Lar+CQ group. Cell invasion was detected by Transwell assay. Scratch test was used to detect cell migration ability. Ki67 immunofluorescence staining was used to detect cell proliferation. Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of epithelialmesenchymal transition related markers in colon cancer cells. Autophagy was detected by adenovirus transfection experiment and transmission electron microscopy. Western blot was used to detect the expression of adenosine monophosphate-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) pathway related proteins. Results Lar significantly inhibited the viability of COLO 205 and HCT 116 cells in a concentration-dependent manner (F=355.181, 403.758, all P<0.001). Compared with the Con group, the number of invasive cells, Ki67 fluorescence intensity and scratch healing rate of colon cancer cells in the Lar group were decreased, the expression of E-cadherin mRNA was increased, the expressions of Vimentin and MMP2 mRNA were decreased, the formation of autophagosomes and autophagic flow, the ratio of microtubule-associated protein light chain 3 (LC3) II/I and p-AMPK/AMPK were increased, and the expression of p62 protein and p-mtor /mTOR ratio were decreased, with significant differences (t= 4.399 ～ 54.214, all P<0.05). Compared with the Lar group, the formation of autophagosome was decreased and the expression of p62 protein was increased in the Lar+CQ group, and the difference was statistically significant (t= 2.755 ～ 24.784, all P<0.05). Conclusion Lar can inhibit the proliferation and migration of colon cancer cells, and the underlying mechanism is related to activation of the AMPK/mTOR signaling pathway and thus inducts autophagy.

参考文献/References:

[1] FABREGAS J C, RAMNARAIGN B, GEORGE T J. Clinical updates for colon cancer care in 2022[J]. Clinical Colorectal Cancer, 2022, 21(3): 198-203.
[2] DEVENPORT S N, SHAH Y M. Functions and implications of autophagy in colon cancer[J]. Cells,2019, 8(11): 1349.
[3] ZHANG Ziqing, LU Yan, ZHANG Hao, et al. Enriched environment ameliorates fear memory impairments induced by sleep deprivation via inhibiting PIEZO1/calpain/autophagy signaling pathway in the basal forebrain[J]. CNS Neuroscience & Therapeutics, 2024,30(2): e14365.
[4] TAVERNARAKIS N. Regulation and roles of autophagy in the brain[J]. Advances in Experimental Medicine and Biology, 2020, 1195: 33.
[5] 宋永嘉, 宋敏, 巩彦龙. 自噬通过BDNF/Trk B 信号通路对脊髓损伤的保护作用[J]. 生命的化学, 2020,40(2): 211-215. SONG Yongjia, SONG Min, GONG Yanlong. Protective effect of autophagy on spinal cord injury through BDNF/Trk B signaling pathway[J]. Chemistry of Life, 2020, 40(2): 211-215.
[6] HONG D S, DUBOIS S G, KUMMAR S, et al. Larotrectinib in patients with TRK fusion-positive solid tumours: a pooled analysis of three phase 1/2 clinical trials[J]. Lancet Oncology, 2020, 21(4): 531-540.
[7] RICCIUTI B, GENOVA C, CRIN? L, et al. Antitumor activity of larotrectinib in tumors harboring NTRK gene fusions: a short review on the current evidence[J]. Onco Targets and Therapy, 2019, 12: 3171-3179.
[8] 罗详冲, 刘晶晶, 李高峰. 拉罗替尼治疗NTRK 基因融合阳性癌症患者的研究进展[J]. 中国肿瘤临床,2019, 46(11): 575-580. LUO Xiangchong, LIU Jingjing, LI Gaofeng. Research progress in the treatment of cancer patients with positive NTRK gene fusion with larotrectinib[J]. Chinese Journal of Clinical Oncology, 2019, 46(11):575-580.
[9] 杨雪晴, 代金连, 吴呈龙, 等. 原肌球蛋白受体激酶小分子抑制剂研究进展[J]. 药学进展, 2023, 47(3):194-206. YANG Xueqing, DAI Jinlian, WU Chenglong, et al. Advances in research on small molecule inhibitors of tropomyosin receptor kinase[J]. Progress in Pharmaceutical Sciences, 2023, 47(3): 194-206.
[10] HAN S Y. TRK inhibitors: Tissue-agnostic anti-cancer drugs[J]. Pharmaceuticals (Basel), 2021, 14(7): 632.
[11] CHEN Yicheng, WANG Huan, CHEN Yuanlei, et al. Trk inhibitor GNF-5837 suppresses the tumor growth,survival and migration of renal cell carcinoma[J]. Oncology Reports, 2019, 42(5): 2039-2048.
[12] 邢增术, 张冲, 刘振湘, 等. siRNA 靶向沉默TrkB基因对人膀胱癌细胞株T24 生长和转移的影响及其分子机制[J]. 中国老年学杂志, 2018, 38(17): 4262-4265. XING Zengshu, ZHANG Chong, LIU Zhenxiang, et al. Effects of siRNA targeting silencing TrkB gene on growth and metastasis of human bladder cancer cell line T24 and its molecular mechanism[J]. Chinese Journal of Gerontology, 2018, 38(17): 4262-4265.
[13] ZOU Jian, ZHANG Zhang, XU Fang, et al. GZD2202,a novel TrkB inhibitor, suppresses BDNF-mediated proliferation and metastasis in neuroblastoma models[J]. Journal of Drug Targeting, 2019, 27(4): 442-450.
[14] ZHANG Yu, HUANG Zhenlin, LI Keqiang, et al. TrkA promotes MDM2-mediated AGPS ubiquitination and degradation to trigger prostate cancer progression[J]. Journal of Experimental & Clinical Cancer Research,2024, 43(1): 16.
[15] KASI P M, AFGHAN M K, BELLIZZI A M, et al. Larotrectinib in mismatch-repair-deficient trk fusionpositive metastatic colon cancer after progression on immunotherapy[J]. Cureus, 2022, 14(7): e26648.
[16] 王希方, 郑伟, 马东瑞, 等. ALKBH5 调控GEFT 的m6A 修饰对胆管癌转移及EMT 的实验研究[J]. 现代检验医学杂志, 2023, 38(4): 1-7, 21. WANG Xifang, ZHENG Wei, MA Dongrui, et al. Experimental study of GEFT m6A modification regulated by ALKBH5 on cholangiocarcinoma metastasis and EMT[J]. Journal of Modern Laboratory Medicine, 2023, 38(4): 1-7, 21.
[17] BUTTACAVOLI M, DI CARA G, ROZ E, et al. Integrated multi-omics investigations of metalloproteinases in colon cancer: focus on MMP2 and MMP9[J]. International Journal of Molecular Sciences, 2021, 22(22): 12389.
[18] 杨琦, 孙正, 朱亦邈, 等. 黄芪- 莪术配伍调控EMT对结肠癌HT-29 细胞增殖?迁移和侵袭能力的影响[J]. 中国中药杂志, 2023, 48(3): 736-743. YANG Qi, SUN Zheng, ZHU Yimiao, et al. Astragali radix-curcumae rhizoma combination inhibits proliferation, migration, and invasion of colon cancer HT-29 cells by regulating EMT[J]. China Journal of Chinese Materia Medica, 2023, 48(3): 736-743.
[19] LEE H J, MOON Y, CHOI J, et al. Characterization of KRC-108 as a TrkA kinase inhibitor with anti-tumor effects[J]. Biomolecules & Therapeutics, 2022, 30(4): 360-367.
[20] VARGAS J N S, HAMASAKI M, KAWABATA T, et al. The mechanisms and roles of selective autophagy in mammals[J]. Nature Reviews Molecular Cell Biology,2023, 24(3): 167-185.
[21] BABAEI G, AZIZ S G, JAGHI N Z Z. EMT, cancer stem cells and autophagy; the three main axes of metastasis[J]. Biomedecine & Pharmacotherapie, 2021,133: 110909.
[22] 韩昭, 高艳玲, 李志锋, 等. 丹参酮Ⅱ A 对雷帕霉素诱导自噬M2 巨噬细胞共培养的皮肤黑素瘤A375细胞EMT 的影响[J]. 皮肤性病诊疗学杂志, 2022,29(4): 291-298. HAN Zhao, GAO Yanling, LI Zhifeng, et al. The mechanism of tanshinone Ⅱ A on regulating EMT in cutaneous melanoma A375 cells through TAMs autophagy[J]. Journal of Diagnosis and Therapy on Dermato-Venereology, 2022, 29(4): 291-298.
[23] WEI Ran, XIAO Yuhong, SONG Yi, et al. FAT4 regulates the EMT and autophagy in colorectal cancer cells in part via the PI3K-AKT signaling axis[J]. Journal of Experimental & Clinical Cancer Research,2019, 38(1): 112.
[24] BENSALEM J, HATTERSLEY K J, HEIN L K, et al. Measurement of autophagic flux in humans: an optimized method for blood samples[J]. Autophagy,2021, 17(10): 3238-3255.
[25] 石拴霞, 阎一鑫, 王纪田, 等. AMPK/mTOR 信号通路在线粒体自噬中的研究进展[J]. 海南医学, 2023,34(5): 736-740. SHI Shuanxia, YAN Yixin, WANG Jitian, et al. Research progress of AMPK/mTOR signaling pathway in mitochondrial autophagy[J]. Hainan Medical Journal,2023, 34(5): 736-740.
[26] HAN Yohan, KEE J Y, HONG S H. Rosmarinic acid activates AMPK to inhibit metastasis of colorectal cancer[J]. Frontiers in Pharmacology, 2018, 9: 68.

相似文献/References:

[1]芦 嘉,史伟峰.血清miRNA-720和miRNA-484在结肠癌分期及早期诊断中的应用[J].现代检验医学杂志,2017,32(01):77.[doi:10.3969/j.issn.1671-7414.2017.01.021]
　LU Jia,SHI Wei-feng.Application of Serum miRNA-720 and miRNA-484 in the Staging and Early Diagnosis of Colon Cancer[J].Journal of Modern Laboratory Medicine,2017,32(06):77.[doi:10.3969/j.issn.1671-7414.2017.01.021]
[2]吴 奇,申高飞,孙丽娜,等.miR-181d在结肠癌中表达失调并抑制癌细胞增殖及凋亡[J].现代检验医学杂志,2017,32(02):15.[doi:10.3969/j.issn.1671-7414.2017.02.004]
　WU Qi,SHEN Gao-fei,SUN Li-na,et al.miR-181d Inhibits Proliferation and Promotes Apoptosis in Colon Cancer Cells[J].Journal of Modern Laboratory Medicine,2017,32(06):15.[doi:10.3969/j.issn.1671-7414.2017.02.004]
[3]李 洪,唐 浏,郑 淋,等.结肠癌组织中microRNA-126表达及临床意义[J].现代检验医学杂志,2017,32(04):60.[doi:10.3969/j.issn.1671-7414.2017.04.017]
　LI Hong,TANG Liu,ZHENG Lin,et al.Expression of microRNA-126 in Colon Cancer and Its Clinical Significance[J].Journal of Modern Laboratory Medicine,2017,32(06):60.[doi:10.3969/j.issn.1671-7414.2017.04.017]

备注/Memo

备注/Memo:
基金项目：陕西省科学技术研究发展计划项目（2020JM4132）。
作者简介：柏希慧（1991-），女，硕士研究生，初级药师，研究方向：临床药理学，E-mail：baixihui5464@126.com。
通讯作者：孙媛媛（1988-），女，本科，主管药师，研究方向：临床药理学，E-mail：348556492@qq.com。
更新日期/Last Update: 2024-11-15