Correlation of SphK1, FAK, Musashi 1 and CA199 expression with angiogenesis and epithelial-mesenchymal transition in surgically removed colon cancer lesions

Objective: To investigate the correlation of SphK1, FAK, Musashi 1 and CA199 expression with angiogenesis and epithelial-mesenchymal transition in surgically removed colon cancer lesions. Methods: A total of 60 patients with colon cancer who underwent radical operation in our hospital between August 2015 and August 2017 were selected, intraoperative colon cancer tissue samples were collected as colon cancer group, and normal tissue specimens adjacent to carcinoma were collected as adjacent tissue group. Fluorescence quantitative PCR was adopted to determine the expression levels of SphK1, FAK, Musashi 1, CA199 as well as the genes related to angiogenesis and epithelial-mesenchymal transition in colon tissues with different properties. Results: SphK1, FAK, Musashi 1 and CA199 mRNA expression in colon cancer group were higher than those in adjacent tissue group;angiogenesis-related genes ANGPTL4, Apelin-13, DLL1, VEGF and HIF-α mRNA expression were higher than those in adjacent tissue group whereas TSP-1 mRNA expression was lower than that in adjacent tissue group;epithelial-mesenchymal transition-related gene E-cadherin mRNA expression was lower than that in adjacent tissue group whereas Vimentin, N-cadherin, Twist and Snail mRNA expression were higher than those in adjacent tissue group. Correlation analysis showed that the SphK1, FAK, Musashi 1 and CA199 expression in colon cancer tissues were directly correlated with the angiogenesis genes and epithelial-mesenchymal transition genes. Conclusion: SphK1, FAK, Musashi 1 and CA199 genes are abnormally expressed in colon cancer tissues and their expression levels are directly correlated with tumor angiogenesis and epithelial-mesenchymal transition process.

Blocking Posttranslational Core Fucosylation Ameliorates Rat Peritoneal Mesothelial Cell Epithelial-Mesenchymal Transition

Background：Core fucosylation （CF）,catalyzed by α-1,6 fucosyltransferase （Fut8） in mammals,plays an important role in pathological processes through posttranslational modification of key signaling receptor proteins,including transforming growth factor （TGF）-β receptors and platelet-derived growth factor （PDGF） receptors.However,its effect on peritoneal fibrosis is unknown.Here,we investigated its influence on epithelial-mesenchymal transition （EMT） of rat peritoneal mesothelial cells （PMCs） in vitro induced by a high-glucose （HG） culture solution.Methods：Rat PMCs were first cultured in a HG （2.5%） culture solution to observe the CF expression level （fluorescein isothiocyanate-lens culinaris agglutinin）,we next established a knockdown model of rat PMCs in vitro with Fut8 small interfering RNA （siRNA） to observe whether inhibiting CF decreases the messenger RNA （mRNA） expression and protein expression of Fut8 and reverses EMT status.Rat PMCs were randomly divided into control group,mock group （transfected with scrambled siRNA）,Fut8 siRNA group,HG group,HG ＋ mock group,and HG ＋ Fut8 siRNA group.Finally,we examined the activation of TGF-β/Smad2/3 signaling and PDGF/extracellular signal-regulated kinase （ERK） signaling to observe the influence of CF on them.Results：CF,Fut8 mRNA,and protein expression were all significantly upregulated in HG-induced EMT model than those in the control rat PMCs （P 〈 0.05）.Fut8 siRNA successfully blocked CF of TGF-β receptors and PDGF receptors and attenuated the EMT status （E-cadherin and α-SMA and phenotypic changes） in HG-induced rat PMCs.In TGF-β/Smad2/3 signaling,Fut8 siRNA did not suppress the protein expression of TGF-3 receptors and Smad2/3;however,it significantly suppressed the phosphowlation of Smad2/3 （relative expression folds of HG ＋ Fut8 group vs.HG group：7.6 ± 0.4 vs.15.1 ± 0.6,respectively,P 〈 0.05）.In PDGF/ERK signaling,Fut8 siRNA did not suppress the protein expression of PDGF receptors and ERK,but it significantly suppressed the phosphorylation of ERK （relative expression folds of HG ＋ Fut8 group vs.HG group：8.7 ± 0.9 vs.15.6 ± 1.2,respectively,P 〈 0.05）.Blocking CF inactivated the activities of TGF-β and PDGF signaling pathways,and subsequently blocked EMT.Conclusions：These results demonstrate that CF contributes to rat PMC EMT.and that blocking it attenuates EMT.CF regulation is a potential therapeutic target of peritoneal fibrosis.

丙泊酚抑制IL-6诱导的A549肺癌细胞上皮间质转化及其机制探讨

目的:探究丙泊酚对白细胞介素-6(IL-6)诱导的A549肺癌细胞上皮间质转化(EMT)的作用及机制。方法:将A549细胞随机分成四组:对照组、IL-6组(50 ng/ml重组IL-6蛋白)、IL-6+丙泊酚低剂量组(50 ng/ml重组IL-6蛋白和5μmol/L丙泊酚)、IL-6+丙泊酚高剂量组(50 ng/ml重组IL-6蛋白和10μmol/L丙泊酚)。MTT法检测细胞活力,Transwell检测细胞迁移情况,Real-time PCR方法检测EMT相关基因(E-cadherin、Vimentin和Snail1)mRNA的表达水平,Western blot检测EMT相关蛋白及JAK2和STAT3的磷酸化表达水平。使用0.5μmol/L STAT3激活剂colivelin处理细胞,检测其对丙泊酚调节的IL-6诱导的A549细胞活力、迁移和EMT的影响。结果:与对照组相比,IL-6组中细胞的活力、迁移、EMT和JAK2/STAT3的活化均增加(P均<0.05);与IL-6组相比,IL-6+丙泊酚组中细胞活力、迁移、EMT和JAK2/STAT3的活化均降低(P均<0.05),这些变化均具有剂量依赖性。STAT3激活剂能够减弱丙泊酚对IL-6诱导的A549细胞活力、迁移和EMT的影响(P均<0.05)。结论:丙泊酚能够抑制IL-6诱导的A549肺癌细胞EMT进程,这种作用是通过抑制JAK2/STAT3的活化发挥作用的。

Salen-Mn通过阻滞TGF-β/Smad2信号通路抑制肾细胞癌786-O细胞的侵袭迁移

目的探讨TGF-β/Smad2信号通路在锰-双希夫碱复合物(Salen-Mananese,Salen-Mn)抑制肾细胞癌侵袭转移中的作用。方法 MTT法检测Salen-Mn对肾细胞癌786-O细胞的细胞活性影响。用伤口愈合实验和Transwell侵袭迁移实验检测Salen-Mn对肾细胞癌786-O细胞的侵袭迁移能力。用Western blot以及real-time PCR检测Salen-Mn对肾细胞癌786-O细胞内上皮间质转化相关蛋白标志物(E-cadherin、N-Cadherin、Vimentin和Snail)的表达。通过Western和siRNA转染技术检测TGF-β/Smad2信号通路在Salen-Mn对EMT调控中的作用。结果 Salen-Mn可以抑制肾细胞癌786-O细胞的增殖和侵袭迁移(P<0.05)。与此同时,Salen-Mn能在mRNA和蛋白水平上调E-cadherin的表达,并下调N-Cadherin、Vimentin和Snail的表达(P<0.05)。Western blot结果显示,Salen-Mn能够抑制肾细胞癌786-O细胞内TGF-β和p-Smad2的表达,且TGF-β的过表达能够逆转Salen-Mn对EMT和侵袭迁移的抑制。结论 Salen-Mn通过下调TGF-β/Smad2信号通路抑制肾细胞癌的侵袭转移,这为肾细胞癌的临床治疗提供了一定的理论依据。

循环肿瘤细胞的异质性在转移性乳腺癌中的作用研究进展

循环肿瘤细胞(CTC)的计数可以预测转移性乳腺癌的预后,但其改善患者预后的能力在临床试验中尚未得到证实。目前研究专注于CTC的分子表征,作为肿瘤组织的"替代物"以非侵入性地方式评估癌症基因组表达及其在治疗过程中的演变。CTC中存在上皮-间质转化过程(EMT),其特点为上皮标志物的缺失。EMT过程可以存在于侵袭性及耐药性较强的细胞,其计数和表征,能够引起肿瘤的复发和进展,具有较高的临床价值。本文深入探讨循环肿瘤细胞的异质性及在转移性乳腺癌上皮-间质转化过程中的作用。使其成为乳腺癌患者监测转移和预后的常规的检测指标,并有助于明确转移的机制,更有望发现乳腺癌转移治疗的新靶点。

吡非尼酮抑制肿瘤相关成纤维细胞促进结肠癌细胞系HT29上皮间质转化的作用及机制

目的:探讨吡非尼酮在肿瘤相关成纤维细胞(CAFs)诱导的HT29结肠癌细胞系侵袭、迁移、增殖、上皮-间质转化(EMT)中的作用及机制。方法:利用共培养小室将CAFs与HT29共培养后CCK-8测定HT29增殖效率;Real time-PCR和Western blot检测HT29结肠癌细胞系E-cadherin和Vimentin转录及蛋白表达水平;Transwell实验检测HT29结肠癌细胞的迁移和侵袭能力。ELISA技术检测CAFs细胞上清液中细胞因子的表达。结果:吡非尼酮抑制CAFs对HT29增殖促进作用(P<0.05);CAFs条件培养基培养HT29后,Vimentin表达水平显著升高(P<0.05),E-cadherin显著降低(P<0.05);吡非尼酮处理CAFs的条件培养基对HT29细胞中两种蛋白无明显影响;Transwell实验表明CAFs与HT29共培养后,其侵袭和迁移能力明显增强(P<0.05);ELISA结果提示CAFs分泌TGF-β1功能受吡非尼酮抑制且存在剂量关系。结论:吡非尼酮通过抑制CAFs分泌TGF-β1抑制CAFs诱导HT29结肠癌细胞系侵袭、迁移、增殖、上皮-间质转化。