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罗哌卡因通过 ITGB1 抑制结直肠癌细胞的增殖和迁移。

Ropivacaine inhibits the proliferation and migration of colorectal cancer cells through ITGB1.

机构信息

Department of Anesthesiology, Beijing Shijitan Hospital, Capital Medical University , Beijing, China.

出版信息

Bioengineered. 2021 Dec;12(1):44-53. doi: 10.1080/21655979.2020.1857120.

Abstract

To study whether ropivacaine inhibits the proliferation and migration of colon cancer cells through ITGB1 (Integrin beta-1). First, the effect of ropivacaine on cell proliferation and migration was detected by MTT and Transwell. DAPI staining, annexin V staining and Western blot were used to detect the expression of apoptosis-related proteins to investigate the effect of ropivacaine on cell apoptosis. Using bioinformatics software to predict the potential drug targets of ropivacaine. RT-PCR, Western blot and immunofluorescence verify the distribution and expression of the drug target ITGB1, and detect its downstream-related proteins to further prove that ropivacaine affects colon cancer cells by acting on ITGB1 protein. 1. Ropivacaine significantly inhibited the proliferation of colon cancer cells and promoted their apoptosis 2. Ropivacaine could interact with ITGB1 protein, and inhibited the expression of ITGB1 protein in colon cancer cells, thereby affecting its downstream signaling pathway. Ropivacaine regulates the function of colon cancer cells by targeting the expression of ITGB1 protein and affecting the activation of its downstream signaling pathways. Integrin beta-1 (ITGB1); 3-(45)-dimethylthiahiazo (-z-y1)-35-di- phenytetrazoliumromide (MTT); 4. 6-diamimo-2-phenyl indole (DAPI); Reverse transcrption PCR (RT-PCR); Colorectal cancer (CRC); Local anesthetics (LA); voltage-gated sodium channel (VGSC); dulbecco s modifed eade medium (DMEM); propidium iodide (PI); dodecyl sulf ate, sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE); Polyvinylidene Fluoride (PVDF); BCL2 associated X (Bax); Focal Adhesion Kinase (FAK); extracellular signal-regulated kmase (ERK); alpha serme threcnime-proteim kinase (AKT); Glyceraldehyde-3-phosphate dehydrogenase (GAPDH); Tris-buffered salme with 0.1% Tween 20 (TBST); Similarty ensemble approach (SEA).

摘要

目的

研究罗哌卡因是否通过 ITGB1(整合素β-1)抑制结肠癌细胞的增殖和迁移。首先,通过 MTT 和 Transwell 检测罗哌卡因对细胞增殖和迁移的影响。使用 DAPI 染色、膜联蛋白 V 染色和 Western blot 检测凋亡相关蛋白的表达,以研究罗哌卡因对细胞凋亡的影响。使用生物信息学软件预测罗哌卡因的潜在药物靶点。RT-PCR、Western blot 和免疫荧光验证药物靶点 ITGB1 的分布和表达,并检测其下游相关蛋白,进一步证明罗哌卡因通过作用于 ITGB1 蛋白影响结肠癌细胞。结果:1. 罗哌卡因显著抑制结肠癌细胞的增殖并促进其凋亡;2. 罗哌卡因可与 ITGB1 蛋白相互作用,并抑制结肠癌细胞中 ITGB1 蛋白的表达,从而影响其下游信号通路。结论:罗哌卡因通过靶向 ITGB1 蛋白的表达并影响其下游信号通路的激活来调节结肠癌细胞的功能。

关键词

整合素β-1(ITGB1);3-(45)-二甲基噻唑 (-z-y1)-35-二苯基四氮唑溴盐(MTT);4.6-二氨基-2-苯基吲哚(DAPI);逆转录聚合酶链反应(RT-PCR);结直肠癌(CRC);局部麻醉剂(LA);电压门控钠通道(VGSC);杜尔贝科改良伊格尔培养基(DMEM);碘化丙啶(PI);十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE);聚偏二氟乙烯(PVDF);B 细胞淋巴瘤 2 相关 X(Bax);粘着斑激酶(FAK);细胞外信号调节激酶(ERK);丝氨酸/苏氨酸蛋白激酶(AKT);甘油醛-3-磷酸脱氢酶(GAPDH);三羟甲基氨基甲烷-盐酸缓冲液与 0.1%吐温 20(TBST)混合;相似性集成方法(SEA)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/268b/8806321/07cc27f9ea1c/KBIE_A_1857120_UF0001_OC.jpg

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