AKT2 S100A4及MMP-9在鼻咽癌组织中的表达

目的:探讨AKT2、S100A4及MMP-9在鼻咽癌组织中的表达、相互关系及临床意义。方法:应用免疫组化EliVision二步法检测50例鼻咽癌组织、25例慢性鼻咽炎组织标本中AKT2、S100A4及MMP-9蛋白的表达,并分析三者与鼻咽癌患者临床特征的关系及三者表达的相关性。结果:鼻咽癌组织中AKT2、S100A4及MMP-9的阳性表达率分别为48.0%(24/50)、52.0%(26/50)、74.0%(37/50),显著高于慢性鼻咽炎组织的8.0%(2/25)、0(0/25)、36.0%(9/25),P均<0.01;AKT2和S100A4的表达均与鼻咽癌颈淋巴结转移及临床分期相关(P均<0.05),MMP-9的表达与鼻咽癌颈淋巴结转移相关(P<0.05),而三者的表达与患者性别、年龄及T分期无关(P均>0.05);鼻咽癌组织中AKT2和S100A4与MMP-9的表达呈正相关(P均<0.01),AKT2与S100A4的表达无关(P>0.05)。结论:AKT2、S100A4及MMP-9的高表达在鼻咽癌的发生、发展及转移方面可具有重要作用,AKT2和S100A4可能通过上调MMP-9的表达而促进鼻咽癌的发展及转移,联合检测三者的表达情况对判断鼻咽癌生物学行为和预测肿瘤转移有一定的临床价值。

S100b protects IMR-32 cells against Ab(1-42) induced neurotoxicity via modulation of apoptotic genes expression

Amyloid beta (1-42) peptide is considered responsible for the formation of senile plaques that accumulate in the brain of patients with Alzheimer’s disease (AD). In the past years considerable attention has been focused on identifying new protective substances that prevent or almost retard the appearance of amyloid beta (1-42)-related neurotoxic effects. In this study, human neuroblastoma cells (IMR-32) was used as system model to evaluate the protective role of S100b, a neurotrophic factor and neuronal survival protein, that is highly expressed by reactive astrocytes in close vicinity of beta-amyloid deposits, against amyloid beta (1-42)-dependent toxicity. Our results show that at nanomolar concentrations, S100b protects cells against Aβmediated cytotoxicity, as assessed by MTS vitality test. The protective mechanism seems to be related to the effect on bcl-2 (an anti-apoptotic gene) expression, which is highly down-regulated by amyloid beta (1-42) treatment, while resulted more expressed in the presence of S100b. On the contrary, Bax, a proapoptotic gene, resulted down-regulated by the treatment with S100 compared with the results obtained in the presence of amyloid beta (1-42) peptide. However, at micromolar doses, S100b is toxic for IMR-32 cells and its toxicity adds to that of the Aβpeptide, suggesting that additional molecular mechanisms may be involved in theneurotoxic process.

Overexpression of S100A4 is closely associated with progression of colorectal cancer

AIM： To investigate whether S100A4 played an important role in the development or progression of colorectal cancer. METHODS： A total of 124 colorectal adenocarcinoma tissue specimens were analyzed by immunohistochemistry for the expression of S100A4 protein and subsequently investigated for the gene mutations in the coding region of S100A4 gene. The specimens were collected over a 3-year period in the laboratories at our large teaching hospital in Seoul, Republic of Korea. RESULTS： Normal colonic epithelium either failed to express or showed focal weak expression of S100A4. Moderate to strong cytoplasmic expression of S100A4 was seen in 69 （55.6%） of the 124 colorectal carcinoma tissue specimens. S100A4 expression was detected in 43 （69.4%） of 62 specimens with lymph node metastasis. Statistically, overexpression of S100A4 was significantly associated with Dukes＇ stage and lymph node metastasis. Nuclear staining was also observed in 24 （19.4%） of 124 samples and closely associated with Dukes＇ stage. However, there was no significant correlation between overexpression of S100A4 and other investigated clinico-pathologic parameters, including tumor localization, tumor size, and survival period. In mutational analysis, no gene mutation was found in the analyzed genomic area of colorectal cancer. CONCLUSION： Overexpression of S100A4 may be closely related with the aggressiveness of colorectal carcinoma.

Prognostic significance of S100A4 and vascular endothelial growth factor expression in pancreatic cancer

AIM: To investigate the expression of vascular endothelial growth factor (VEGF) and calcium-binding protein S100A4 in pancreatic cancer and their relationship to the clinicopathological parameters and prognosis of pancreatic cancer. METHODS: Expression status of VEGF and S100A4 was examined in 62 surgical specimens of primary pancreatic cancer by immunohistochemistry. Correlation between the expression of VEGF and S100A4 and clinicopathological parameters was analyzed. RESULTS: Thirty-eight of 62 (61.3%) specimens of primary pancreatic cancer were positive for S100A4. Thirty-seven (59.7%) specimens showed positive expression of VEGF. The positive correlation between S100A4 and VEGF expression was significant in cancer tissues (P < 0.001). S100A4 expression was significantly correlated with tumor size, TNM stage and poorer prognosis. VEGF expression had a significant correlation with poorer prognosis. The prognosis of 17 S100A4-and VEGF-negative cancer patients was significantly better than that of other patients (P < 0.05). Distant metastasis (P = 0.001), S100A4-(P = 0.008) and VEGF-positive expression (P = 0.016) were significantly independent prognostic predictors (P < 0.05). CONCLUSION: Over-expression of S100A4 and VEGF plays an important role in the development of pancreatic cancer. Combined examination of the two molecules might be useful in evaluating the outcome of patients with pancreatic cancer.

Expression of protein S100A4 is a predictor of recurrence in colorectal cancer

AIM: To investigate the prognostic significance of S100A4 expression in colorectal cancer and its correlation with expression of E-cadherin and p53. METHODS: A cohort of archival formalin-fixed paraff inembedded specimens was selected from 127 patients with colorectal cancer who underwent surgical resection between April 2000 and March 2004 at the Department of Surgery, Korea University Guro Hospital. The expression of protein S100A4 was evaluated according to the proportion of positively stained cancer cells. In each case, three core biopsies with a diameter of 2 mm were punched out and positioned in a recipient paraff in array block. Four-μm sections of these tissue array blocks were used for immunohistochemical analysis of protein S100A4, E-cadherin, and p53. Clinicopathological data were based on the original histopathologic reports and clinical records of patients. RESULTS: In normal colorectal mucosa, protein S100A4 immunoreactivity was clearly absent in both cytoplasm and nucleus. However, positive immunoreactivity of protein S100A4 was detected in 45 (35.4%) of the tumor cases. There was no significant association between positive immunoreactivity of protein S100A4 and clinicopathological parameters such as tumor differentiation or TNM stage, and also no correlation between the reactivity and E-cadherin or p53 expression. However, positive immunoreactivity of protein S100A4 was found to be associated with tumor recurrence (P = 0.004), and was also associated with significantly worse overall survival in the Kaplan-Meyer survival analysis (P = 0.044). After adjustment for tumor differentiation, tumor depth and nodal status, however, it failed to achieve statistical significance (P = 0.067). CONCLUSION: The expression of protein S100A4 is associated with tumor recurrence and poor overall survival in patients with colorectal cancer.

Significance of KiSS-1 and S100A4 in the process of metastasis of gastric carcinoma

Objective： To detect the expression of KISS-1 and S100A4 in the primary tumor tissues and lymphatic and visceral metastases and investigate its role in tumorigenesis and metastasis of gastric cancer. Methods： The protein expression of KISS-1 and S100A4 in lymphatic and visceral metastases from advanced gastric cancer specimens was mainly examined by immunohistochemical staining and tissues microarray. Results： Immunohistochemical staining revealed reduced expression of KISS-1 and up-regulated expression of S100A4 in lymph node and visceral metastases. Rates of KISS-1 expression in normal tissues, primary tumor tissues, lymph node and visceral metastases were 95.8%, 74.6%, 60.9% and 57.5%. $100A4 expression in associated cases was 43.6%, 71.8%, 70.3% and 90.0%, respectively. Significant differences in KISS-1 expression was significantly higher in normal tissues than that in primary tumor tissues （P〈0.001）. While significant differences of S100A4 expression could be seen between normal and cancer tissues （P〈0.001） and between visceral and primary tumors （P〈0.05）. Conclusion： Tumor metastasis results from gradual accumulation of abnormal genetic alterations. Down-regulation of KISS-1 and up-regulation of S100A4 play a critical role in metastasis of gastric carcinoma.

妊娠糖尿病患者血清S100A4、FNDC5水平及其与妊娠结局的关系

目的 探究妊娠糖尿病(GDM)患者血清S100钙结合蛋白A4(S100A4)、Ⅲ型纤连蛋白结构域包含蛋白5(FNDC5)水平及其与妊娠结局的关系。方法 选取2020年6月至2022年6月该院126例GDM患者作为GDM组,选取同期孕检的126例健康孕妇作为对照组。根据GDM患者妊娠结局情况分为妊娠结局良好组(76例)和妊娠结局不良组(50例)。采用酶联免疫吸附试验(ELISA)检测血清S100A4、FNDC5水平,采用Logistic回归分析GDM孕妇妊娠结局,采用受试者工作特征(ROC)曲线评估血清S100A4、FNDC5预测GDM孕妇妊娠不良结局的价值,采用Spearman相关性分析血清S100A4、FNDC5与不良妊娠结局的相关性。结果 GDM组血清S100A4水平明显高于对照组(P<0.05),血清FNDC5水平明显低于对照组(P<0.05)。妊娠结局良好组血清S100A4水平明显低于妊娠结局不良组(P<0.05),血清FNDC5水平明显高于妊娠结局不良组(P<0.05)。Spearman相关性分析结果显示,GDM患者血清S100A4水平与不良妊娠结局呈正相关(P<0.05),FNDC5水平与不良妊娠结局呈负相关(P<0.05)。Logistic回归分析表明,血清S100A4、空腹血糖、HOMA-IR是GDM患者发生不良妊娠结局的危险因素(P<0.05),FNDC5是GDM患者发生不良妊娠结局的保护因素(P<0.05)。血清S100A4、FNDC5联合预测GDM妊娠结局的曲线下面积大于S100A4(Z=2.045,P=0.041)及FNDC5单项检测(Z=2.010,P=0.044)。结论 GDM患者血清S100A4水平升高,FNDC5水平降低,二者对评估GDM孕妇妊娠结局具有一定的参考价值。

重型颅脑损伤患者血清S100A4、S100A12检测的临床意义分析

目的探讨重型颅脑损伤(sTBI)患者血清S100钙结合蛋白(S100)A4、S100A12检测的临床意义。方法选择2021年3月至2023年3月邯郸市中心医院收治的147例sTBI患者作为sTBI组,另选取同期于邯郸市中心医院体检的体检健康者50例作为对照组。分析S100A4、S100A12水平与脑损伤标志物的相关性和sTBI早期死亡的影响因素,并研究血清S100A4、S100A12对sTBI早期死亡的预测价值。结果sTBI组血清S100A4、S100A12、髓鞘碱蛋白(MBP)、S100B、神经元特异性烯醇化酶(NSE)水平均明显高于对照组(P<0.05);sTBI患者血清S100A4、S100A12水平与MBP、NSE、S100B均呈正相关(P<0.05);多因素Logistic回归分析结果显示,入院时格拉斯哥昏迷量表(GCS)评分降低,血清S100A4、S100A12、MBP、NSE、S100B水平升高为sTBI患者早期死亡的危险因素(P<0.05);受试者工作特征曲线结果显示,血清S100A4、S100A12联合入院时GCS评分、MBP、NSE、S100B预测sTBI患者早期死亡的效能优于单项检测。结论sTBI患者血清中S100A4和S100A12水平升高,其与脑损伤加重和发生早期死亡有关,S100A4联合S100A12检测对于sTBI早期死亡具有较好的预测价值。

METTL14 downregulation drives S100A4^(+)monocyte-derived macrophages via MyD88/NF-κB pathway to promote MAFLD progression

Without intervention,a considerable proportion of patients with metabolism‐associated fatty liver disease(MAFLD)will progress from simple steatosis to metabolism‐associated steatohepatitis(MASH),liver fibrosis,and even hepatocellular carcinoma.However,the molecular mechanisms that control progressive MAFLD have yet to be fully determined.Here,we unraveled that the expression of the N6-methyladenosine(m6A)methyltransferase METTL14 is remarkably downregulated in the livers of both patients and several murine models of MAFLD,whereas hepatocyte-specific depletion of this methyltransferase aggravated lipid accumulation,liver injury,and fibrosis.Conversely,hepatic Mettl14 overexpression alleviated the above pathophysiological changes in mice fed on a high-fat diet(HFD).Notably,in vivo and in vitro mechanistic studies indicated that METTL14 downregulation decreased the level of GLS2 by affecting the translation efficiency mediated by YTHDF1 in an m6A-depedent manner,which might help to form an oxidative stress microenvironment and accordingly recruit Cx3cr1^(+)Ccr2^(+)monocyte-derived macrophages(Mo-macs).In detail,Cx3cr1^(+)Ccr2^(+)Mo-macs can be categorized into M1-like macrophages and S100A4-positive macrophages and then further activate hepatic stellate cells(HSCs)to promote liver fibrosis.Further experiments revealed that CX3CR1 can activate the transcription of S100A4 via CX3CR1/MyD88/NF-κB signaling pathway in Cx3cr1^(+)Ccr2^(+)Mo-macs.Restoration of METTL14 or GLS2,or interfering with this signal transduction pathway such as inhibiting MyD88 could ameliorate liver injuries and fibrosis.Taken together,these findings indicate potential therapies for the treatment of MAFLD progression.

钙结合蛋白S100A4对BCG感染THP-1细胞自噬的调控作用

本研究旨在探究牛结核分枝杆菌减毒株卡介苗(Bacillus Calmette-Guérin,BCG)感染人单核巨噬细胞THP-1后,钙结合蛋白S100A4对细胞自噬的调控作用。以感染复数为10∶1的BCG感染THP-1细胞不同时间,用Western blot检测S100A4和LC3Ⅱ的表达,以确定最佳感染时间,并采用透射电镜观察自噬体数量变化。在BCG单独感染或与S100A4小干扰RNA共处理THP-1细胞12 h后,采用qRT-PCR检测S100A4及自噬相关因子--微管相关蛋白轻链3II (microtubule-associated protein light chain 3Ⅱ, LC3Ⅱ)、自噬相关蛋白7(autophagy-related gene 7, Atg7)、Beclin-1在mRNA水平上的表达,采用Western blot检测S100A4、LC3Ⅱ、Atg7、Beclin-1在蛋白水平的表达。利用mRFP-GFP-LC3检测自噬流,激光共聚焦显微镜观察细胞中mRFP-LC3和mRFP-GFP-LC3点状聚集。结果显示:在BCG感染THP-1细胞不同时间后,S100A4和LC3Ⅱ在蛋白表达水平随感染时间的延长先上升后下降,在12 h时表达最高(P<0.001),且自噬体数量明显增多。在mRNA水平上,与siNC组相比,siNC+BCG感染组S100A4、LC3Ⅱ、Atg7、Beclin-1的mRNA表达水平显著上调(P<0.05),当BCG和siS100A4共处理后,S100A4、LC3Ⅱ、Atg7、Beclin-1在mRNA水平的表达显著(P<0.05)或极显著(P<0.01,P<0.001)下调;在蛋白水平上,与未感染组相比,S100A4、LC3Ⅱ、Atg7、Beclin-1的蛋白表达量显著增多(P<0.05),BCG和siS100A4共处理后,S100A4、LC3Ⅱ、Atg7、Beclin-1的蛋白表达量均极显著减少(P<0.001);与未感染组相比,BCG组自噬体的数量极显著增多(P<0.01),siS100A4+BCG组与siNC+BCG组相比,自噬体的数量显著减少(P<0.05)。S100A4对BCG感染巨噬细胞诱导的自噬具有调控作用,S100A4能够促进BCG诱导的THP-1细胞自噬。

结膜黑色素瘤组织中S100A4蛋白表达及其预后预测价值

目的探究结膜黑色素瘤组织中S100A4蛋白表达及其预后预测价值。方法选取2021年1月至2023年1月于灵宝市福寿堂眼科医院、郑州大学第一附属医院接受手术治疗的56例结膜黑色素瘤患者为研究对象。采用免疫组化法S-P法检测S100A4蛋白在结膜黑色素瘤组织中的表达,根据检测结果将入组患者分为S100A4蛋白高表达组(n=35)和S100A4蛋白低表达组(n=21),比较S100A4表达与总生存时间的关系。依据结膜黑色素瘤患者的预后情况将入组患者分为预后良好组(n=18)、预后不良组(n=38),分析可能影响结膜黑色素瘤患者预后不良的危险因素,根据危险因素构建Logistic回归预测模型,采用受试者工作特征曲线评估该预测模型效能。结果结膜黑色素瘤患者中位总生存时间为10个月,术后1 a存活率为80.36%(45/56)。S100A4蛋白高表达组患者中位生存时间、术后1 a生存率(9个月,74.58%)均低于S100A4蛋白低表达组(10个月,90.47%;Log-rankχ^(2)=3.485,P=0.001)。预后不良组患者肿瘤低分化、淋巴结转移、S100A4蛋白高表达比例分别为68.42%、73.68%、71.05%,均高于预后良好组的22.22%、27.78%、22.22%(χ^(2)=10.481,P=0.001;χ^(2)=10.635,P=0.001;χ^(2)=11.785,P=0.001)。多因素Logistic回归分析结果显示,肿瘤低分化(OR=2.042,95%CI:1.350~2.734)、淋巴结转移(OR=1.876,95%CI:1.225~2.526)、S100A4蛋白高表达(OR=1.828,95%CI:1.354~2.301)是影响黑色素瘤患者预后的危险因素。S100A4蛋白表达Logistic回归预测模型预测结膜黑色素瘤患者预后的敏感性、准确性、阳性预测值、曲线下面积分别为92.16%、90.02%、89.34%、0.914。结论肿瘤低分化、淋巴结转移、S100A4蛋白高表达是影响结膜黑色素瘤患者预后不良的危险因素,基于上述因素构建的模型预测结膜黑色素瘤患者预后不良的效能良好。

糖尿病肾病患者血清GDF-11、S100A4水平与病情、疾病转归的关系分析

目的探讨血清生长分化因子-11(GDF-11)、S100钙结合蛋白A4(S100A4)水平与糖尿病肾病(DN)患者病情严重程度、疾病转归的关系。方法选取2021年5月至2023年1月该院收治的95例DN患者作为研究组,另选取110例体检健康者作为健康组。将DN患者根据病情严重程度分为轻度组(n=66)、重度组(n=29),患者出院后连续随访半年,根据预后情况分为预后良好组(n=64)和预后不良组(n=31)。采用酶联免疫吸附试验检测血清GDF-11、S100A4水平,采用Spearman等级相关分析探讨血清GDF-11、S100A4水平与病情严重程度的关系,采用受试者工作特征(ROC)曲线评估血清GDF-11、S100A4对DN患者疾病转归的预测价值,采用多因素Logistic回归分析DN患者疾病转归的影响因素。结果轻度组、重度组血清GDF-11、S100A4水平高于健康组,且重度组高于轻度组(P<0.05)。DN患者血清GDF-11、S100A4水平与病情严重程度均呈正相关(P<0.05)。预后良好组血清GDF-11、S100A4水平低于预后不良组(P<0.05)。血清GDF-11、S100A4预测DN患者疾病转归的曲线下面积(AUC)分别为0.785、0.839,二者联合预测的AUC为0.902。预后不良组2型糖尿病(T2DM)病程、肾小球分级为Ⅲ~Ⅳ级、间质炎症评分为2分、间质性纤维化和小管萎缩(IFTA)评分为2~3分、估算肾小球滤过率<60 mL/(min·1.73 m^(2))占比及总胆固醇、甘油三酯、低密度脂蛋白胆固醇、24 h尿蛋白定量、糖化血红蛋白、C肽、血细胞比容、红细胞沉降率水平高于预后良好组(P<0.05)。多因素Logistic回归分析显示,T2DM病程≥12.0年、IFTA评分≥2分、GDF-11≥700.82 ng/mL、S100A4≥211.53 ng/L是DN患者预后不良的危险因素(P<0.05)。结论血清S100A4、GDF-11水平在DN患者中呈高表达,且与病情严重程度及疾病转归相关,有望作为评估DN病情及预后的潜在标志物。

S100A4在银屑病炎症中的机制研究

目的:探究S100钙结合蛋白A4 (S100 Calcium Binding Protein A4, S100A4)在银屑病炎症中的作用机制。方法:在HaCat细胞中使用S100A4的抗体进行改进的紫外交联免疫共沉淀结合高通量测序(improved RNA Binding Protein Immunoprecipitation high throughput sequencing, iRIP-seq)技术获得与S100A4互作的RNA,将相关基因序列比对KEGG数据库来明确S100A4调控的炎症通路。结果:S100A4结合的靶RNA所在基因与MAPK信号通路、核糖体、内质网中的蛋白质加工、糖尿病并发症中的AGE-RAGE信号通路、膀胱癌、慢性髓样白血病、弓形体病、结肠直肠癌、破骨细胞分化、军团杆菌病等功能通路的调节有关,表明S100A4可能是调节银屑病及其并发症的一种关键炎症介质。结论:S100A4在银屑病炎症机制中具有潜在的调控作用,这些发现为S100A4作为银屑病发病炎症介质提供了新的证据。

S100A4在肥大细胞的表达及血清S100A4含量的检测对过敏性哮喘的诊断价值

目的初步研究S100钙结合蛋白A4(S100A4)是否参与肥大细胞脂质代谢以及对过敏性哮喘的临床意义。方法从基因表达数据集(GEO)数据库中下载过敏性哮喘相关的单细胞RNA(scRNA)表达矩阵GSE213085,使用R包进行数据可视化处理。通过体外培养C57肥大细胞,慢病毒感染肥大细胞建立S100A4低表达的细胞系,实时荧光定量PCR和Western blot法分别检测S100A4的mRNA和蛋白水平,实时荧光定量PCR检测脂质代谢相关基因ATP柠檬酸合成酶(ACLY)、乙酰辅酶A羧化酶1(ACC1)、CD36、溶质载体家族27a成员6(SCL27a6)、过氧化物酶体增殖物激活受体α(PPARα)的mRNA水平。采集过敏性哮喘的患者和健康对照人群外周血,ELISA检测血清S100A4水平,绘制受试者工作特征(ROC)曲线,分析S100A4对疾病的诊断效能,并计算曲线下面积(AUC)。结果用scRNA数据集GSE213085鉴定出26225个细胞,提取出肥大细胞进行后续分析,功能富集分析结果表明,肥大细胞特征基因集中于脂质代谢相关的途径。肥大细胞低表达S100A4时,其脂质代谢相关基因ACLY、ACC1、CD36、SCL27a6、PPARα表达下调。ELISA检测显示过敏性哮喘患者血清S100A4较健康对照人群升高,ROC曲线发现,血清S100A4含量对过敏性哮喘具有一定诊断效能(AUC=0.746)。结论S100A4可能参与肥大细胞脂质代谢调节,血清S100A4检测对过敏性哮喘诊断可能有一定价值。

钙结合蛋白S100A4通过TLR4/NF-κB信号通路调控胶质瘤细胞增殖、存活及迁移功能

目的:探索外源性钙结合蛋白S100A4对胶质瘤细胞增殖、存活及迁移功能的影响及机制。方法:利用UCSC数据库下载泛癌数据集,进行S100A4在泛癌中的表达及预后分析;从CGGA数据库中下载胶质瘤患者的转录组数据和临床数据,利用R软件进行分析S100A4表达在胶质瘤患者中的预后与进展;利用在线分析工具GEPIA与STRING进行胶质瘤患者的S100A4蛋白质网络互作分析。体外培养人胶质瘤细胞系(U87与U251),实验分为3组:PBS组、S100A4组、S100A4+TAK242组[瑞沙托维(TAK242)为Toll-样受体4(TLR4)特异性抑制剂]。流式细胞术检测胶质瘤细胞的增殖与凋亡;细胞划痕实验、Transwell实验及克隆形成实验检测U87与U251细胞的迁移及增殖能力;Western blot检测TLR4蛋白、NF-κB相关信号分子蛋白水平。结果:生物信息学结果显示,S100A4在多种肿瘤中显著上调(P<0.05),包括:胶质母细胞瘤(GBM)、低级胶质瘤(LGG)、胃癌(STAD)、肝细胞肝癌(LIHC)等,其中在GBM和LGG中预后较差。与低表达S1004的胶质瘤患者相比,高表达S100A4的胶质瘤患者生存期更短,WHO分级程度更高。此外,胶质瘤患者S100A4蛋白可能与膜联蛋白A2(ANXA2)、TLR4和晚期糖基化终末产物特异性受体(AGER)蛋白存在相互作用。细胞实验中,与PBS组相比,外源性S100A4处理后,胶质瘤U87和U251细胞增殖、迁移能力增强,TLR4、p-ERK1/2、p-p38、p-p65蛋白水平显著上调(P<0.05)。与S100A4组相比,S100A4+TAK242组胶质瘤细胞增殖、迁移能力减弱,TLR4、p-ERK1/2、p-p38、p-p65蛋白水平显著下调(P<0.05)。结论:S100A4可能通过TLR-4/NF-κB信号通路调控胶质瘤细胞增殖、迁移及存活功能。

S100A4核定位在胰腺癌转移中的作用与机制研究

目的:研究S100A4核定位在胰腺癌(pancreatic cancer,PC)转移中的作用与机制。方法:利用PC病程进展的组织芯片,通过免疫组化和HALO数字病理精准分析平台等技术,定量S100A4在组织细胞,特别是胞核中的表达情况,统计分析各检测指标与临床参数及生存期之间的关系。通过分子结构信息学分析、质粒构建与转染以构建基因调控的不同分组细胞研究模型;利用核质蛋白分离、免疫共沉淀、Western blot、划痕实验、Transwell实验、靶向剪切及转座酶技术(cleavage under targets and tagmentation,CUT&Tag)等研究S100A4核定位在PC转移中的作用与机制。结果:S100A4的高表达及其核定位与PC的T、N分期和不良预后正相关。PC细胞中S100A4核定位受小泛素相关修饰物(small ubiquitin-related modifier,SUMO)修饰调控,泛素结合酶9介导了PC细胞中S100A4的K22和K96位点与SUMO1结合,并通过SUMO特异性蛋白酶1实现去SUMO化,动态平衡PC细胞中S100A4的SUMO化修饰水平;调控S100A4蛋白的SUMO化修饰可改变PC细胞的体外转移能力;CUT&Tag测序结果证明S100A4核定位参与调控与肿瘤转移功能相关的基因网络。结论:S100A4核定位可提示PC预后不良,有望成为临床治疗方案制定的重要依据。发现阻断或抑制S100A4核定位的办法,可能成为抑制PC转移,特别是早期转移,改善PC患者预后的新靶点。

原发性肾病综合征患儿血清LXA4、S100A4水平与病情、疾病转归的关系

目的探讨原发性肾病综合征(PNS)患儿血清脂氧素A4(LXA4)、S100钙结合蛋白A4(S100A4)水平与病情、疾病转归的关系。方法选取2020年3月至2022年3月南京明基医院收治的168例PNS患儿为研究对象,另选取来南京明基医院体检的150例健康儿童为对照组,将PNS患儿根据疾病活动度分为缓解期组(n=73)与发作期组(n=95)。采用酶联免疫吸附试验检测血清LXA4、S100A4水平。患儿出院后连续随访1年,根据预后情况分为预后良好组(n=138)与预后不良组(n=30)。采用Spearman相关分析血清LXA4、S100A4水平与疾病活动度的关系,采用受试者工作特征(ROC)曲线评估血清LXA4、S100A4对PNS患儿预后的预测价值;采用多因素Logistic回归分析PNS患儿预后的影响因素。结果缓解期组、发作期组血清LXA4、S100A4水平高于对照组,且发作期组血清LXA4、S100A4水平高于缓解期组(P<0.05)。PNS患儿血清LXA4、S100A4水平与疾病活动度均呈正相关(r=0.593、0.546,P<0.05)。预后良好组PNS患儿血清LXA4、S100A4水平低于预后不良组(P<0.05)。血清LXA4、S100A4预测PNS患儿疾病转归的曲线下面积(95%CI)分别为0.767(0.756~0.888)、0.846(0.824~0.863),二者联合预测的曲线下面积(95%CI)为0.905(0.879~0.924)。预后不良组病理类型为肾炎型占比、24 h尿蛋白定量、血尿发生占比高于预后良好组,尿蛋白转阴时间长于预后良好组,病理类型为单纯型及总蛋白、IgM水平低于预后良好组(P<0.05)。多因素Logistic回归分析显示,有血尿发生(OR=2.300,95%CI 1.598~3.312)、24 h尿蛋白定量≥121.25 mg/24 h(OR=2.098,95%CI 1.489~2.956)、LXA4≥95.64 ng/L(OR=2.627,95%CI 1.737~3.973)、S100A4≥248.15 ng/L(OR=2.729,95%CI 1.777~4.192)是PNS患儿预后不良的危险因素(P<0.05)。结论血清S100A4、LXA4水平在PNS患儿中升高,且与疾病活动度及预后相关,有望作为评估PNS病情及预后的潜在标志物。

胃肠道间质瘤组织S100A4、Slingshot-1表达与临床病理特征及预后关系

目的:探讨胃肠道间质瘤组织S100钙结合蛋白A4(S100A4)、弹弓酶-1(Slingshot-1)表达情况及与患者临床病理特征及预后的关系。方法:2010年1月至2014年6月,胃肠道间质瘤患者115例切除的瘤组织和瘤旁正常组织样本,采用实时定量PCR(RT-PCR)检测组织样本中S100A4、Slingshot-1的表达,分析其与临床病理特征的关系,Kaplan-Meier生存曲线评估S100A4、Slingshot-1表达与患者术后5年总生存率的关系,Cox回归模型分析胃肠道间质瘤患者预后的影响因素。结果:胃肠道间质瘤组织S100A4、Slingshot-1相对表达量均高于瘤旁正常组织(P均<0.05);胃肠道间质瘤组织S100A4表达与肿瘤直径、浸润深度、NIH危险度、远处转移有关(P均<0.05),Slingshot-1表达与浸润深度、NIH危险度、远处转移有关(P均<0.05)。Kaplan-Meier生存分析表明,S100A4高表达者、Slingshot-14高表达者的术后5年总生存率均分别低于S100A4低表达者、Slingshot-14低表达者(P<0.05)。Cox回归模型多因素分析结果表明,肿瘤直径、NIH危险度、远处转移、S100A4高表达、Slingshot-14高表达是影响胃肠道间质瘤患者预后的危险因素(P均<0.05)。结论:胃肠道间质瘤组织S100A4、Slingshot-1表达增加,与患者病情进展和不良预后有关,监测S100A4、Slingshot-1表达可能有利于患者的预后判断。

S100A4基因载体的构建及其在人胃癌细胞系中的表达

目的 克隆S100A4 cDNA,构建pcDNA3.1-S100A4重组表达载体,转染胃癌细胞系MKN1,观察S100A4在胃癌细胞中的表达情况。方法 Trizol法提取人胃上皮细胞GES-1的总RNA,逆转录反应获得含S100A4基因的cDNA。聚合酶链反应GenBank获得S100A4基因序列,并设计引物,利用聚合酶链反应(PCR)扩增出分子量为327 bp的产物。构建pMD18-T simple-S100A4重组质粒,转化JM109菌。菌液测序成功后,提取质粒,进行BamHⅠ/HindⅢ双酶切,酶切产物回收纯化,连接表达载体pcDNA3.1,构建pcDNA3.1-S100A4真核表达载体。利用脂质体介导转染方法将纯化的表达载体转染到胃癌细胞系MKN1中,采用逆转录聚合酶链反应(RT-PCR)检测转染后MKN1细胞中S100A4 mRNA表达水平。结果 PCR产物连接克隆载体的测序结果与GenBank公布的无突变序列一致,HindⅢ/BamHⅠ双酶切可以成功构建真核表达载体pcDNA3.1-S100A4;在胃癌细胞系MKN1中转染pcDNA3.1-S100A4表达载体,以转染pcDNA3.1空载体和正常未转染的MKN1细胞作为对照。结果表明,转染pcDNA3.1-S100A4表达载体48 h后,S100A4 mRNA表达水平升高,差异有统计学意义(P<0.01)。结论 构建完成pcDNA3.1-S100A4真核表达载体,S100A4基因在胃癌细胞MKN1中成功表达。

S100A4蛋白与过敏性疾病的关系研究进展

S100A4属于S100蛋白家族,在成纤维细胞、炎性细胞和巨噬细胞中均广泛表达[1-3],通过晚期糖基化终产物受体(RAGE)和toll样受体4(TLR4)参与调节多种细胞的生长、存活、分化与运动[4-8]。目前,S100A4的临床研究主要集中在癌症转移促进特性[9]。