胆管细胞性肝癌组织CSNK...与肿瘤转移和血管生成的关系_唐津天.pdf

东南大学学报(医学版)J Southeast Univ(Med Sci Edi)2023，Feb;42(1):40-48Ophthalmol Vis Sci，2016，57(10):4356-4366 18ZHENG H J，ZHANG X，GUO J，et al Lysosomal dysfunction-induced autophagic stress in diabetic kidney diseaseJ JCell Mol Med，2020，24(15):8276-8290 19CHE H，LI H，LI Y，et al Melatonin exerts neuroprotectiveeffects by inhibiting neuronal pyroptosis and autophagy inSTZ-induced diabetic miceJ FASEB J，2020，34(10):14042-14054 20ZHANG L G，LI P L，DAI Y，et al Mibefradil alleviates high-glucose-induced cardiac hypertrophy by inhibiting PI3K/Akt/mTO-mediated autophagyJ J Cardiovasc Pharma-col，2020，76(2):246-254 21TANIDA I Autophagosome formation and molecular mecha-nism of autophagyJ Antioxid edox Signal，2011，14(11):2201-2214 22SHIN W H，PAK J H，CHUANG K C The central regulatorp62 between ubiquitin proteasome system and autophagy andits role in the mitophagy and Parkinson s diseaseJ BMBep，2020，53(1):56-63 23 DIETICH J B Apoptosis and anti-apoptosis genes in theBcl-2 familyJ Arch Physiol Biochem，1997，105(2):125-135 24BENTNALL M，ODIGUEZ-MENOCAL L，DE GUEVA-A L，et al Caspase-9，caspase-3 and caspase-7 have dis-tinct roles during intrinsic apoptosisJ BMC Cell Biol，2013，14(71):32-41 收稿日期2022-07-07修回日期2022-11-10 基金项目新疆维吾尔自治区自然科学基金资助项目(2021D01C399)作者简介唐津天(1981 )，男，新疆乌鲁木齐人，副主任医师，医学博士。E-mail:lili03021163 com 引文格式唐津天，唐润娟，薛峰，等 胆管细胞性肝癌组织 CSNK11 的表达及其与肿瘤转移和血管生成的关系J 东南大学学报(医学版)，2023，42(1):40-48 论著 胆管细胞性肝癌组织 CSNK11 的表达及其与肿瘤转移和血管生成的关系唐津天1，唐润娟2，薛峰1，黎旺红1(1 新疆医科大学附属肿瘤医院 肝胆胰外科二病区，新疆 乌鲁木齐830000;2 新疆医科大学第二附属医院康复科，新疆 乌鲁木齐830000)摘要目的:探究酪蛋白激酶 11(caseinkinase1alpha1，CSNK11)在胆管细胞性肝癌(intrahepatic cholan-giocarcinoma，ICC)组织中的表达，及其对肿瘤细胞转移及血管生成的影响。方法:收集新疆医科大学附属肿瘤医院 50 例 ICC 组织及癌旁组织样本，实时荧光定量 PC(qT-PC)法和免疫组化染色法检测 CSNK11表达水平;将 ICC 细胞系 HUCCT-1 培养后随机分为对照组、si-NC 组、si-CSNK11 组，通过脂质体介导法将靶向 CSNK11 基因的短发夹 NA 序列转染至 HUCCT-1 细胞，qT-PC 和蛋白质免疫印迹法(Western blot-ting)测定细胞内 CSNK11mNA 与蛋白的表达变化，以鉴定转染效果;细胞划痕实验和 Transwell 实验检测细胞的迁移与侵袭能力变化，细胞免疫荧光染色观察细胞内上皮间质转化标志物钙黏附蛋白 E(E-cadherin)和波形蛋白(Vimentin)的表达强度，血管生成实验检测肿瘤血管生成情况，Western blotting 检测细胞中血管内皮生长因子(vascular endothelial growth factor，VEGF)、基质金属蛋白酶(matrix metallopeptidase，MMP)2(MMP-2)及 MMP-9 的蛋白表达变化。结果:ICC 组织 CSNK11 mNA 相对表达量显著高于癌旁组织，且ICC 组织 CSNK11 阳性率也显著高于癌旁组织(P 0 01);与对照组和 si-NC 组比较，si-CSNK11 组 HUC-CT-1 细胞中 CSNK11 mNA 相对表达量和蛋白相对表达量均下调，细胞划痕愈合率降低，细胞迁移与侵袭数目均减少，E-cadherin 荧光密度值下降而 Vimentin 荧光密度值升高，细胞管样形成数目减少，细胞内04VEGF、MMP-2 及 MMP-9 的蛋白相对表达量均显著下调，差异均具有统计学意义(P 0 05)。结论:CSNK11 在 ICC 组织内呈高表达，靶向下调 ICC 细胞 CSNK11 表达后可抑制细胞迁移、侵袭及上皮间质转化过程，并减少血管生成，从而控制肿瘤进展。关键词胆管细胞性肝癌;酪蛋白激酶 11;转移;血管生成 中图分类号735 7 文献标志码A 文章编号1671-6264(2023)01-0040-09doi:10 3969/j issn 1671-6264 2023 01 006Expression of CSNK11 in cholangiocarcinoma and its relationshipwith tumor metastasis and angiogenesisTANG Jintian1，TANG unjuan2，XUE Feng1，LI Wanghong1(1 The Second Ward of Hepatobiliary Pancreatic Surgery，Affiliated Tumor Hospital of Xinjiang Medical University，Urumqi 830000，China;2 Department of ehabilitation，the Second Affiliated Hospital of Xinjiang Medical University，Urumqi 830000，China)AbstractObjective:To investigate the expression of casein kinase 1 alpha1(CSNK11)in intrahepatic cholan-giocarcinoma(ICC)tissues and its effect on tumor cell metastasis and angiogenesis Methods:Fifty ICC tissuesamples and adjacent tissue samples from Affiliated Tumor Hospital of Xinjiang Medical University were collected，and the expression level of CSNK11 was detected by real-time quantitative PC(qT-PC)and immunohisto-chemical staining The ICC cell line HUCCT-1 was cultured and randomly divided into control group，si-NC groupand si-CSNK11 group The short hairpin NA sequence targeting CSNK11 gene was transfected into HUCCT-1cells by liposome-mediated method，the expression changes of CSNK11 mNA and protein in cells were measuredby qT-PC and Western blotting to identify the transfection effect Cell scratch assay and Transwell assay wereused to detect changes in cell migration and invasion abilities，cell immunofluorescence staining was used to observethe expression intensity of intracellular epithelial-mesenchymal transition markers E-cadherin and Vimentin，angio-genesis assays was used to detect tumor angiogenesis，the protein expression changes of vascular endothelial growthfactor(VEGF)，matrix metallopeptidase(MMP)2(MMP-2)and MMP-9 in cells were detected by Western blot-ting esults:The relative expression of CSNK11 mNA in ICC tissue was significantly higher than that in adja-cent tissue，and the positive rate of CSNK11 in ICC tissue was also significantly higher than that in adjacent tissue(P 0 01)Compared with the control group and si-NC group，the relative expression of CSNK11 mNA andprotein in HUCCT-1 cells of si-CSNK11 group were down-regulated，the wound healing rate of cells decreased，the number of cell migration and invasion decreased，the fluorescence density value of cadherin decreased while thefluorescence density value of Vimentin increased，and the number of cell tube-like formation decreased，at the sametime，the relative proteins expressions of VEGF，MMP-2 and MMP-9 in cells were significantly d