Downregulation of MUC1 Inhibits Proliferation and Promotes Apoptosis by Inactivating NF-κB Signaling Pathway in Human Nasopharyngeal Carcinoma

Objective To investigate the effect of mucin 1(MUC1)on the proliferation and apoptosis of nasopharyngeal carcinoma(NPC)and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital.The expression of MUC1 was measured by real-time quantitative PCR(qPCR)in the patients with PNC.The 5-8F and HNE1 cells were transfected with siRNA control(si-control)or siRNA targeting MUC1(si-MUC1).Cell proliferation was analyzed by cell counting kit-8 and colony formation assay,and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells.The qPCR and ELISA were executed to analyze the levels of TNF-αand IL-6.Western blot was performed to measure the expression of MUC1,NFкB and apoptosis-related proteins(Bax and Bcl-2).Results The expression of MUC1 was up-regulated in the NPC tissues,and NPC patients with the high MUC1 expression were inclined to EBV infection,growth and metastasis of NPC.Loss of MUC1 restrained malignant features,including the proliferation and apoptosis,downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells.Conclusion Downregulation of MUC1 restrained biological characteristics of malignancy,including cell proliferation and apoptosis,by inactivating NF-κB signaling pathway in NPC.

Mucin 1 and interleukin-11 protein expression and inflammatory reactions in the intestinal mucosa of necrotizing enterocolitis children after surgery

BACKGROUND Necrotizing enterocolitis(NEC)of the newborn is a frequently occurring clinical disease in infants.The mortality rate of NEC in premature infants is as high as 50%,and the morbidity rate is on the rise.NEC has already caused serious impacts on newborn survival and poses serious threats to both children and families.AIM To investigate the expression and significance of mucin 1(MUC1)and interleukin-11(IL-11)in the intestinal mucosa of infants with neonatal NEC after surgery.METHODS Forty-eight postoperative intestinal mucosal specimens from children with NEC(NEC group)and twenty-two intestinal mucosal specimens from children with congenital intestinal atresia(control group)were collected in our hospital.Immunohistochemical staining and Western blot analysis were used to examine the protein expression of MUC-1 and IL-11 in the two groups.The serum levels of tumor necrosis factor-α(TNF-α)and IL-1βin the two groups were measured by enzyme-linked immunosorbent assay,and the relationship between MUC-1 and IL-11 protein expression and serum TNF-αand IL-1βlevels was analyzed by the linear correlation method.RESULTS The protein expression of MUC-1 and IL-11 in the NEC group was significantly lower than that in the control group,and the difference was statistically significant(P<0.05).The levels of serum TNF-αand IL-1βin the NEC group were significantly higher than those in the control group(P<0.05).The protein expression of MUC-1 and IL-11 in the NEC group negatively correlated with serum TNF-αand IL-1βlevels(P<0.05).There was a significant negative correlation between the protein expression of MUC-1 and IL-11 and the levels of serum TNF-αand IL-1βin the NEC group.CONCLUSION The protein expression of MUC1 and IL-11 in the intestinal mucosa of children with NEC is significantly downregulated after surgery.This downregulation may be involved in the pathogenesis of this disease and has a certain correlation with inflammatory response factors in children with NEC.

Immunohistochemical molecular markers as predictors of curability of endoscopically resected submucosal colorectal cancer

AIM: To clarify the usefulness of immunohistochemical molecular markers in predicting lymph node metastasis of submucosal colorectal cancer. METHODS: We examined microvessel density, lymphatic vessel density, the Ki-67 labeling index, expression of MUC1 and Matrix metalloproteinase-7 (MMP-7) in tumor cells, and expression of cathepsin D in stromal cells at the invasive front by immunostaining of samples resected from 214 patients with submucosal colorectal cancer. Pathologic features were assessed on hematoxylin-eosin- stained samples. We evaluated the relations between clinicopathologic/immunohistochemical features and lymph node metastasis. RESULTS: Lesions of the superficial type, with an unfavorable histologic grade, budding, lymphatic involvement, high microvessel density (≥ 40), high lymphatic vessel density (≥ 9), high Ki-67 labeling index (≥ 42), and positivity of MUC1, cathepsin D, and MMP-7 showed a significantly high incidence of lymph node metastasis. Multivariate analysis revealed that high microvessel density, unfavorable histologic grade, cathepsin D positivity, high lymphatic vessel density, superficial type, budding, and MUC1 positivity were independent risk factors for lymph node metastasis.A combined examination with four independent immunohistochemical markers (microvessel density, cathepsin D, lymphatic vessel density, and MUC1) revealed that all lesions that were negative for all markers or positive for only one marker were negative for lymph node metastasis. CONCLUSION: Analysis of a combination of immuno- histochemical molecular markers in endoscopically resected specimens of submucosal colorectal cancer allows prediction of curability regardless of the pathologic features visible of hematoxylin-eosin-stained sections.

Clinicopathological utility of sialoglycoconjugates in diagnosing and treating colorectal cancer

Aberrant expression of glycoconjugates occurs during malignant transformation of cancer cells.Overexpression of sialoglycoconjugates in particular may play an important role in the progression,i.e.,invasion or metastasis,of cancer.Various types of sialoglycoconjugates have been investigated to clarify their biological significance and clinical utility in diagnosing and treating colorectal cancer.This review focuses specifically on expression of mucin(MUC)1 and it suggests that MUC1with the specific structure of a sialo-oligosaccharide has biological significance in determining the metastatic potential of colorectal cancer cells and clinicopathological utility in evaluating the effectiveness of treatments and the prognosis for patients with colorectal cancer.Further studies are expected to contribute to the expanded use of cancer-associated sialoglycoconjugates in cancer diagnosis and therapy.

Invasive ductal carcinoma of the pancreas showing exophytic growth

BACKGROUND:Invasive pancreatic carcinoma generally appears as poorly defined mass reflecting the infiltrative growth.We aimed to identify the histological and immunohistochemical features in a rare case of pancreatic carcinoma showing exophytic growth. METHODS:A 67-year-old woman presented with a mass of 5.0 cm in diameter in the pancreatic head.Preoperative computed tomography revealed a well-demarcated, primarily solid mass with a central low-density area. Magnetic resonance cholangiopancreatography revealed neither encasement nor dilation of the main pancreatic duct.An incorrect preoperative diagnosis was made of solid pseudopapillary tumor of the pancreas.Elevated serum carcinoembryonic antigen(CEA)levels and abnormal FDG positron emmission tomography accumulation suggested that the tumor had malignant potential requiring a pancreatoduodenectomy. RESULTS:The head of the pancreas contained a well- circumscribed encapsulated mass of 5.0 cm in diameter, comprising 50%adenocarcinoma,with mucinous carci- noma in the center and anaplastic carcinoma at the periphery.The anaplastic carcinoma comprised pleo- morphic cells(PCs)and pleomorphic giant cells(PGCs). The PGCs phagocytozed mononuclear PCs and lymphocytes adjacent to the capsule without infiltrating the capsule itself.Immunohistochemistry revealed that the anaplastic carcinoma cells including PGCs were positive for the tumor antigen Mucin 1 and CEA but negative for vimentin. CONCLUSION:Our observations suggest anaplastic carcinoma components in the present tumor have a ductal origin and that the exophytic tumor growth is associated with the phagocytotic activity of PGCs.

多态性上皮黏蛋白1和原癌基因C-myc在老年甲状腺乳头状癌患者中的表达

目的 探讨多态性上皮黏蛋白1 （mucin 1,MUC1）和原癌基因蛋白质（proto-oncogene proteins C-myc,C-Myc）基因在老年甲状腺乳头状癌（PTC）患者中的表达和临床意义. 方法 应用免疫组化法检测30例腺瘤旁正常甲状腺组织、35例结节性甲状腺肿和58例PTC标本中MUC1和C-myc的表达水平. 结果 MUC1基因在甲状腺癌中表达率为77.6％（45/58）,MUC1在有局部浸润和淋巴结转移患者中的检出率分别为90.0％（9/10）和88.2％（15/17）.C-myc基因在甲状腺癌中阳性率为81.0％ （47/58）;C-myc在淋巴结转移组中的检出率为100.0％ （17/17）. 结论 MUC1和C myc的阳性表达在甲状腺良恶性病变间有差异,并与甲状腺癌的转移有关.

CircHECTD1 up-regulates mucin 1 expression to accelerate hepatocellular carcinoma development by targeting microRNA-485-5p via a competing endogenous RNA mechanism

Background::Non-coding RNAs have attracted considerable attention for their vital role in cancer.The purpose of this study was to determine the effects of non-coding RNAs on hepatocellular carcinoma(HCC)and reveal their regulatory mechanism in the pathophysiological process.Methods::We measured the expression of mucin 1(MUC1)and miR-485-5p in tissues from 15 HCC patients and in liver cancer cell lines by quantitative real-time polymerase chain reaction and Western blot,screened for aberrantly expressed microRNAs(miRNAs)by miRNA microarrays.Bioinformatics tools were used to find the miRNA and circular RNA that regulated MUC1,which were validated by RNA immunoprecipitation assay and luciferase reporter assay.Cell counting kit-8,Transwell assays,and flow cytometry were used to conduct functional experiments.Proteins were examined by western blot and immunohistochemical staining assays.Significant differences between groups were estimated using the one-way analysis of variance.A P<0.05 was considered statistically significant.Results::MUC1 was overexpressed in HCC tissues compared with that in paratumor tissues(normal vs.tumor,1.007±0.215 vs.75.213±18.403,t=18.401,P<0.001)while miR-485-5p was down-regulated(normal vs.tumor,4.894±0.684 vs.1.586±0.398,t=16.191,P<0.001).Inhibition of miR-485-5p promoted cell proliferation(73.33%±5.13%vs.41.33%±3.51%,t=8.913,P<0.001),migration(102±8 cells vs.46±8 cells,t=8.681,P<0.001),invasion(59±7 cells vs.28±2 cells,t=8.034,P<0.01),and suppressed apoptosis(22.64%±6.97%vs.36.33%±3.96%,t=2.958,P<0.05)of HepG2 cells with which MUC1 is knocked down.Mechanically,miR-485-5p binds to MUC1,while circHECTD1 binds to miR-485-5p,resulting in the indirect up-regulation of the MUC1 level.Conclusions::Our findings reveal that circHECTD1 facilitates HCC progression by sponging miR-485-5p to up-regulate MUC1.

黏蛋白1与多发性骨髓瘤免疫治疗的研究进展

多发性骨髓瘤（MM）是一种目前不可被治愈的恶性浆细胞肿瘤。免疫治疗在消除MM微小残留病、降低复发、提高患者总生存率方面有重要作用,受到人们越来越多的重视。黏蛋白1（MUC1）为MM相关抗原，以其为靶点的免疫治疗具有良好的应用前景，其中部分MUC1疫苗已经进入临床试验，并报道有临床反应。该文讨论MUC1及其作为MM免疫治疗靶点的最新研究进展。

Fine Particulate Matter-Induced Exacerbation of Allergic Asthma via Activation of T-cell Immunoglobulin and Mucin Domain 1

Background： Fine particulate matter （PM2.5） exacerbates airway inflammation and hyperreactivity in patients with asthma, but the mechanism remains unclear. The aim of this study was to observe the effects of prolonged exposure to high concentrations of PM2.5 on the pathology and airway hyperresponsiveness （AHR） of BALB/c mice undergoing sensitization and challenge with ovalbumin （OVA） and to observe the effects of apoptosis and T-cell immunoglobulin and mucin domain 1 （TIM-1） in this process. Methods： Forty female BALB/c mice were divided into four groups： control group, OVA group, OVA/PM group, and PM group （n - 10 in each group）. Mice in the control group were exposed to filtered clean air. Mice in the OVA group were sensitized and challenged with OVA. Mice in the OVA/PM group were sensitized and challenged as in the OVA group and then exposed to PM2.5 for 4 h per day and 5 days per week for a total of 8 weeks using a nose-only ＂PM2.5 online enrichment system＂ in The Second Hospital of Hebei Medical University. Mice in the PM group were exposed to the PM2.5 online enrichment system only. AHR was detected. Bronchoalveolar lavage fluid （BALF） was collected for cell classification. The levels of interleukin-4 （IL-4）, IL-5, and IL-33 in BALF were measured using enzyme-linked immunosorbent assay. Changes in histological structures were examined by light microscopy, and changes in ultramicrostructures were detected by electron microscopy. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL） assay in the lung tissues. Western blotting and immunohistochemistry were utilized to analyze the expression of Bcl-2, Bax, and TIM-1 in the lungs. Results： The results showed that AHR in the OVA/PM group was significantly more severe than that in the OVA and PM groups （P 〈 0.05）. AHR in the PM group was also considerably more severe than that in the control group （P 〈 0.05）. The BALF of OVA/PM group （28.00± 6.08 vs. 12.33 ±4.51, t = 4.631, P = 0.002） and PM group （29.00 ± 3.00 vs. 12.33 ± 4.51, t = 4.927, P = 0.001） had more lymphocytes than the BALF of the control group. The number of neutrophils in the BALF of the OVA/PM group （6.67 ± 1.53 vs. 3.33 ± 1.53, t = 2.886, P = 0.020） and PM group （6.67 ± 1.53 vs. 3.33 ± 1.53, t = 2.886, P = 0.020） was much higher than those in the BALF of OVA group （P 〈 0.05）. TUNEL assays showed that the number of apoptotic cells in the OVA/PM group was significantly higher than that in the OVA group （Tunel immunohistochemical scores [IHS%], 1.20 ± 0.18 vs. 0.51 ± 0.03, t = 8.094, P 〈 0.001） and PM group （Tunel IHS%, 1.20 ± 0.18 vs. 0.51 ±0.09, t = 8.094, P〈 0.001）, and that the number of apoptotic cells in the PM group was significantly higher than that in the control group （Tunel IHS%, 0.51 ± 0.09 vs. 0.26 ± 0.03, t = 2.894, P = 0.020）. The concentrations of IL-4 （77.44 ± 11.19 vs. 48.02 ±10.02 pg/ml, t = 4.595, P= 0.002） and IL-5 （15.65 ± 1.19vs. 12.35±0.95pg/ml, t=3.806,P=0.005） and the Bax/Bcl-2 ratio （1.51 ± 0.18 vs. 0.48 ± 0.10, t = 9.654, P 〈 0.001） and TIM-1/β-actin ratio （0.78 ± 0.11 vs. 0.40 ±0.06, t = 6.818, P 〈 0.001） in the OVA/PM group were increased compared to those in the OVA group. The concentrations of IL-4 （77.44 ± 11.19 vs. 41.47 ± 3.40 pg/ml, t = 5.617, P= 0.001） and IL-5 （ 15.65±1. 19 vs. 10.99 ± 1.40 pg/ml, t = 5.374, P = 0.001 ） and the Bax/Bcl-2 ratio （1.51 ±0.18 vs. 0.97 ± 0.16, t = 5.000, P = 0.001） and TIM-1/β-actin ratio （0.78 ± 0.11 vs. 0.31 ± 0.06,t = 8.545, P 〈 0.001 ） in the OVA/PM group were increased compared to those in the PM group. The concentration oflL-4 （41.47 ±3.40 vs. 25.46 ± 2.98 pg/ml, t = 2.501, P = 0.037） and the Bax/Bcl-2 ratio （0.97 ± 0.16 vs. 0.18 ± 0.03, t = 7.439, P 〈 0.001） and TIM-1/β-actin ratio （0.31 ± 0.06 vs. 0.02 ± 0.01, t = 5.109, P = 0.001） in the PM group were also higher than those in the control group. Conclusions： Exacerbated AHR associated with allergic asthma caused by PMz5 is related to increased apoptosis and TIM-1 activation. These data might provide insights into therapeutic targets for the treatment of acute exacerbations of asthma induced by PM2.5.

慢病毒介导的人MUC1核心肽串联重复片段修饰的树突状细胞疫苗的制备

目的探讨制备慢病毒介导的人mucin糖粘蛋白1(MUC1)核心肽串联重复序列(TR)修饰的树突状细胞(DC)疫苗的可行性。方法人工合成编码MUC1 5个TR片段的基因序列,将其克隆入慢病毒载体pLV-GFP中,经酶切、基因测序鉴定。构建成功的慢病毒载体转染293T细胞,制备病毒。病毒液感染DC细胞。流式细胞术检测感染效率及DC表型,RT-PCR及Westernblot检测MUC1目的基因的表达。结果酶切及测序证实慢病毒载体构建成功。制备的慢病毒能有效感染DC细胞,感染效率达32.54%;流式检测证实慢病毒感染后DC成熟表型未受影响;RT-PCR及Western blot检测证实转染后的DC可有效表达MUC1。结论成功制备慢病毒介导的MUC1基因TR片段修饰的DC疫苗。

Glycopeptide ligation via direct aminolysis of selenoester

Direct aminolysis of selenoester in aqueous media was investigated as a glycopeptide ligation strategy.This strategy allows the peptide and glycopeptide ligation to proceed smoothly(even with hindered amino acids) without the need of cysteine residue, N-terminal thiol auxiliary or coupling additive, and to afford the corresponding amide products in excellent yields. No epimerization was observed during ligation reations. In this work, the selenoester of unprotected glycopeptide was readily prepared, and the direct aminolysis of glycopeptide selenoester was successfully applied to synthesize MUC1 mucin sequence efficiently.