High number of intraepithelial CD8+ tumor-infiltrating lymphocytes is associated with the absence of lymph node metastases in patients with large early-stage cervical cancer

In a prospective study, we have examined the tumor-specific immune response in a group of 59 patients with human papillomavirus (HPV) 16-positive (HPV16(+))-induced or HPV18(+)-induced cervical cancer. Local antitumor immunity was analyzed by the enumeration of tumor-infiltrating dendritic cells and CD4+, CD8+, and regulatory T cells as well as by calculation of the ratio of CD8+/CD4+ T cells and CD8+/regulatory T cells. Systemic tumor-specific immunity was assessed by determination of the HPV E6- and/or E7-specific T-cell response in the blood of these patients. Finally, these variables were evaluated with respect to known histopathologic prognostic variables, including the absence (LN-) or presence (LN+) of lymph node metastases. Stratification according to the lymph node status of patients revealed a significantly stronger CD8+ T-cell tumor infiltration, a higher CD8+/CD4+ T-cell ratio, and higher CD8+/regulatory T-cell ratio in the group of patients in which the tumor failed to metastasize to the tumor-draining lymph node. Subdivision according to the presence (IR+) or absence (IR-) of circulating HPV-specific T cells disclosed that the highest number of tumor-infiltrating CD8+ T cells was found in the group of LN- patients displaying a concomitant systemic tumor-specific immune response (LN-IR+). CD8+ T-cell infiltration in LN-IR- patients was comparable with that of LN+ patients. In cervical cancer, the absence of lymph node metastases is strongly associated with a better prognosis. Our data indicate that, especially in a subgroup of LN- patients, a strong and effective interaction between immune system and tumor exists. This subgroup of cervical cancer patients may have the best prognosis.     

乳腺癌患者淋巴结CD4＋CD25＋T细胞和相关细胞因子的检测及相关性分析

目的探讨CD4＋CD25＋T细胞在乳腺癌发生、发展中的作用。方法将取自35例乳腺癌患者的105个肿大淋巴结,制备成单细胞悬液,应用流式细胞仪检测CD4＋CD25＋T细胞比例及CD4＋CD25-、CD8＋T细胞、NK细胞的相对水平。采用定量RT-PCR法,检测IL-2、IL-10、TGF-β1和IFN-γ的细胞因子水平。结果乳腺癌患者淋巴结中的CD4＋CD25＋T细胞水平（在CD4＋T细胞中的百分含量）与淋巴结转移相关,转移淋巴结中其水平明显高于未转移淋巴结。乳腺癌患者淋巴结中CD4＋CD25＋T细胞与CD4＋CD25-、CD8＋T细胞和NK细胞的水平呈负相关关系。乳腺癌患者淋巴结中CD4＋CD25＋T细胞水平与TGF-β1呈正相关,与IL-2、IL-10、IFN-γ无相关性。乳腺癌患者淋巴结中TGF-β1、IL-10、IFN-γ水平与淋巴结转移相关,转移淋巴结中TGF-β1、IL-10含量高而IFN-γ含量较低。IL-2与淋巴结转移无相关性。结论乳腺癌患者转移淋巴结中的CD4＋CD25＋T细胞水平高于未转移淋巴结。     

肝细胞性肝癌组织浸润淋巴细胞表型与分布研究

目的:肝细胞性肝癌组织浸润淋巴细胞与外周血T 细胞表型可能与肿瘤进展及预后相关,本研究检测肝癌患者组织及外周血T 细胞表型与分布,分析淋巴细胞表型变化与预后的关系。方法:分析2007年10月至12月中山医院147 例肝癌及癌旁组织浸润淋巴细胞表型（T 细胞或B 细胞表面标志物:CD3、CD8、CD4、CD20、CD19、Foxp 3）,表型与临床病理特征及预后的关系;检测26例肝癌外周血CD3、CD8、CD4 +T细胞数量并其比例变化。结果:癌巢内肿瘤浸润细胞明显少于癌周组织（P < 0.01）,癌周淋巴细胞主要分布于癌旁正常肝组织、汇管区,其与患者肝炎病史及肝硬化相关,表型以CD3 +T细胞为主,其中又以CD8 + 细胞毒性T 细胞为主;CD4 染色在多数病例为阴性,Foxp 3 仅在个别病例（15/ 109）呈阳性。肿瘤浸润淋巴细胞B 细胞标志CD20、CD19均为阴性。肿瘤组织内CD8 +T细胞浸润数量与预后正相关,而癌周浸润淋巴细胞数目与患者转移及复发无显著关系。结论:肝癌肿瘤浸润细胞在癌巢内明显少于癌周组织,肿瘤及癌周浸润细胞以CD8 + 细胞毒性T 细胞为主。肿瘤组织内CD8 +T细胞浸润数量与预后相关,而癌周浸润淋巴细胞数量与患者转移及复发无显著关系。      

A novel role of metalloproteinase in cancer-mediated immunosuppression

Depressed immune responses have been observed frequently in cancer patients. In a variety of human malignancies, the expression of interleukin-2 receptor alpha (IL-2R alpha) on activated tumor-infiltrating lymphocytes was down-regulated. Because IL-2R alpha plays a pivotal role in the development and propagation of functional T cells, its depressed expression may result in poor function of tumor-reactive cytotoxic lymphocytes. For elucidating the mechanism responsible for down-regulation of IL-2R alpha, a coculture model of in vitro mixed autologous lymphocytes and tumor cells was established. Kinetic analysis showed that cervical cancer cells down-regulated IL-2R alpha expression on encountered T cells. The amount of IL-2R alpha mRNA in tumor-infiltrating lymphocytes-derived CD8+ T cells was compatible with that in the corresponding activated CD8+ T cells. Additional evidence showed that cervical cancer cells could induce the release of soluble IL-2R alpha expression on encountered T cells. By using protease inhibition assays we demonstrated that tissue inhibitors of metalloproteinase abrogated the cancer-mediated IL-2R alpha proteolytic process and restored the T-cell proliferation function. Immunohistochemical stainings further revealed prominent metalloproteinase (MMP) expressions, including MMP-1, MMP-2, and MMP-9, in cervical cancer tissues. Additional in vitro studies showed that MMP-9 mediates cleavage of IL-2R alpha and down-regulates the proliferative capability of cancer-encountered T cells. Our findings suggest a new role of MMPs in tumor-mediated immunosuppression and provide a possible therapeutic potential for patients with cervical cancer.     

Reversed CD4/CD8 ratios of tumor-infiltrating lymphocytes are correlated with the progression of human cervical carcinoma.

BACKGROUND: To investigate the clinical significance of tumor-infiltrating lymphocytes (TILs) within the tumor milieu of human cervical carcinoma, the authors quantitatively measured and compared the subpopulations of lymphocytes infiltrating the neoplastic cervix. METHODS: A total of 30 patients with Stage Ia-IIa cervical carcinoma were enrolled. TILs were isolated from tissue specimens by means of a mechanical dispersal technique, and the immunocyte subsets were quantified with dual-color flow cytometry. Bulky tumor was defined as tumor size >4 cm in greatest dimension according to the 1995 staging of the International Federation of Gynecology and Obstetrics. RESULTS: The CD4/CD8 ratios of TILs were reversed in both cervical squamous cell carcinoma (n = 20) and cervical adenocarcinoma (n = 10). The proportion of CD4(+) T cells was significantly lower in tumors from patients with lymph node metastasis (n = 8) than in those from patients without lymph node metastasis (n = 22) (24.5 vs. 32.7, P = 0.001), as was the reversed CD4/CD8 ratio (0.50 vs. 0.81, P = 0.001). The proportion of CD4(+) T cells was much lower in bulky tumors (n = 5) than in nonbulky tumors (n = 25) (21.4 vs. 32.5, P < 0.001), reflecting in a more strongly reversed CD4/CD8 ratio (0.41 vs. 0.81, P = 0.001). CONCLUSIONS: Decreased proportions of tumor-infiltrating CD4+ T cells with reversed CD4/CD8 ratios are highly correlated with rapid tumor growth and lymph node metastasis in cervical carcinoma. The regional immune escape is of prognostic importance with regard to cancer progression.     

COX-2、CD44V6与卵巢癌恶性生物学指标的关系

目的 探讨环氧化酶-2(COX-2)、CD44变异性V6(CD44V6)在卵巢癌组织中的表达与肿瘤恶性生物学指标的关系.方法 选取90例卵巢癌标本(卵巢癌组)、45例卵巢良性肿瘤组织标本(良性组),检测2组标本中的COX-2蛋白、CD44V6蛋白表达,并分析不同临床分期、分化程度、淋巴结转移情况下的肿瘤组织中的COX-...     

三阴性乳腺癌组织中PD-L1和CD8+ TIL的水平及其临床意义

目的：探讨程序性死亡蛋白-配体 1（programmed death ligand-1,PD-L1）和肿瘤浸润淋巴细胞（tumor-infiltrating lymphocyte, TIL）在三阴性乳腺癌（triple-negative breast cancer,TNBC）组织中的水平及其临床意义。方法：收集2015年1月至 2019年1月福建医科大学附属第二医院手术切除的 61 例 TNBC 患者的癌及癌旁组织石蜡标本,用免疫组化法检测癌组织中 PD-L1表达和CD8+ TIL的水平,用卡方检测方法分析TNBC 组织中PD-L1 和 CD8+ TIL 水平与患者临床病理特征及预后的关 系。结果：PD-L1和CD8+TIL在TNBC组织中的阳性率分别为63.9%（39/61）和32.8%（20/61）。PD-L1表达与TNBC患者的肿瘤 大小、淋巴结转移、病理分期、复发与否有明显关联（均P<0.05）,与患者的年龄、肿瘤分化程度、脉管侵犯以及Ki67表达水平无明 显关联（均P>0.05）;CD8+ TIL水平与TNBC 患者的肿瘤大小、肿瘤分化程度、淋巴结转移、病理分期、复发与否有明显关联 （均P<0.05）,与患者的年龄、脉管侵犯以及Ki67表达水平无明显关联（均P>0.05）。PD-L1和CD8+TIL水平与患者的无进展生存 期（PFS）及总生存期（OS）具有显著相关性（均P<0.05）,PD-L1+ 或者缺乏CD8+ TIL与患者更差的PFS及OS相关（均P<0.05）。结 论：TNBC组织中存在较高水平的PD-L1和CD8+TIL,PD-L1阳性表达或缺乏CD8+TIL与肿瘤侵袭性增加相关,也与患者更差的 PFS及OS相关。     

L1CAM在甲状腺乳头状癌组织及细胞中的表达及与临床病理特征的关系

目的:检测甲状腺乳头状癌组织及细胞株中L1CAM表达,探讨其与临床病理特征的相关性及意义。方法:选取手术切除甲状腺乳头状癌组织、对应的癌旁组织及正常甲状腺组织标本各32例,甲状腺癌细胞株及正常甲状腺细胞株。采用Western blot及qRT-PCR检测组织及细胞株中L1CAM的表达,观察其与肿瘤部位、直径、数目、分化程度、淋巴结转移及AJCC分期等临床病理参数的关系。结果:L1CAM蛋白及mRNA在甲状腺乳头状癌组织中表达高于癌旁甲状腺组织,在正常甲状腺组织无表达（P<0.05）;甲状腺乳头状癌组织中L1CAM蛋白水平与肿瘤直径、肿瘤数目、淋巴结转移、AJCC分期具有相关性（P均＜0.05）,肿瘤直径大于1 cm、肿瘤多发、淋巴结转移及AJCC分期Ⅲ-Ⅳ的甲状腺癌组织中L1CAM表达量高;L1CAM蛋白及mRNA在BHT101、B-CPAP甲状腺癌细胞株中均有表达,在正常甲状腺上皮细胞NTHY-ORI 3-1无表达,在BHT101中高于其他细胞株（P<0.05）。结论:L1CAM在甲状腺乳头状癌组织及细胞中表达上调,与甲状腺乳头状癌发展及转移恶性临床病理特征相关。     

CXCL12-CXCR4生物学轴与甲状腺乳头状癌淋巴结转移的相关性研究

探讨CXCL12-CXCR4生物学轴与甲状腺乳头状癌淋巴结转移的相关性。方法:采用半定量RT-PCR和免疫组织化学法分别检测72例新鲜甲状腺乳头状癌及淋巴结组织,52例甲状腺乳头状癌及淋巴结石蜡组织中CXCR4、CXCL12 mRNA及蛋白的表达。结果:甲状腺乳头状癌组织及转移灶组织中CXCR4 mRNA及蛋白高表达,淋巴结转移组的表达显著高于非转移组,差异有统计学意义（P<0.05）;颈部淋巴结组织中CXCL12 mRNA及蛋白均高表达,转移淋巴结及非转移淋巴结差异无统计学意义（P>0.05）。结论:CXCR4、CXCL12的表达与甲状腺乳头状癌淋巴结转移密切相关,推测CXCL12-CXCR4生物学轴在甲状腺乳头状癌转移的过程中发挥重要作用,CXCR4可作为抑制甲状腺乳头状癌转移的有效靶点。      

肺腺癌浸润CD8+T细胞表达CD39的机制

目的：探讨肿瘤浸润CD8⁺T细胞表达CD39的可能机制。方法：通过TCGA数据库的肺腺癌(LUAD)组织及正常肺组织转录组数据分析CD39在LUAD组织和正常肺组织中的表达差异及其对患者预后的影响,分析CD39表达与T细胞浸润、激活的关系。用小鼠LUAD Lewis细胞建立小鼠皮下移植瘤模型,FCM检测淋巴结、脾脏以及移植瘤组织中CD8⁺CD39⁺T细胞。收集Lewis细胞培养上清液作为条件培养基(CCM),免疫磁珠法(MACS)分选CD8⁺T细胞、CD11b⁺细胞;在培养基中分别加入CCM、IL-6和采取非接触或接触培养方式进行培养,探索CD8⁺T细胞表达CD39的可能机制。结果：CD39在LUAD组织中呈低表达(P<0.01),其表达水平与LUAD患者OS、T细胞浸润和激活水平均呈正相关(P<0.05或P<0.001)。FCM检测结果显示,在移植瘤组织中CD8+CD39⁺T细胞的比例明显高于淋巴结及脾脏(P<...     

Immunohistochemical study of leukocyte infiltration and expression of hsp70 in esophageal squamous cell carcinoma

It is reported that macrophages and CD4+ or CD8+ cytotoxic T cells have an important role in the suppression of cancer progression. The aim of this study was to clarify these immune responses in patients with esophageal cancer. We enrolled 28 patients with pT2 esophageal cancer that had been resected without preoperative adjuvant therapy. The correlations between the numbers of infiltrating CD4+, CD8+ and CD68+ cells, the expression of heat shock protein 70 (hsp70) and a variety of clinicopathologic factors were analyzed. The numbers of CD8+ T cells and CD68+ macrophages showed a significant positive correlation with tumor diameter (p = 0.01, p = 0.037) and the expression of hsp70 (p = 0.01, p = 0.02) and a negative correlation with lymph node metastasis (p = 0.0079, p < 0.0001). The expression of hsp70 exhibited a negative correlation with lymph node metastasis (p = 0.023). CD8+ T cells and CD68+ macrophages might have a suppressive function against esophageal cancer progression. Our results suggested that hsp70 might play an important role in the presentation of tumor specific antigens.     

BMPR2 expression level is correlated with low immune infiltration and predicts metastasis and poor survival in osteosarcoma

Osteosarcoma is the most common malignant bone tumor in adolescents and young adults, and identifying biomarkers for prognosis and therapy is necessary. Bone morphogenetic protein receptor 2 (BMPR2) is involved in various cellular functions, including cell adhesion, proliferation and invasion, inflammation, apoptosis and metastatic spread. However, the correlation between BMPR2 expression levels and prognosis and tumor-infiltrating immune cells in osteosarcoma is not well understood. In the present study, the expression level of BMPR2 was investigated using the Oncomine and R2 databases. The association between the expression level of BMPR2 and the clinical prognosis of patients with cancer was analyzed using the R2 database. The relationship between the expression level of BMPR2 and immune cell infiltration in the stroma of osteosarcoma was assessed using the Tumor Immune Estimation Resource (TIMER) and CIBERSORT. The correlations between BMPR2 expression level and infiltrated immune cell gene marker sets in osteosarcoma were validated in the TIMER and R2 databases. Analysis of a cohort of patients with osteosarcoma revealed that BMPR2 expression was significantly higher in osteosarcoma compared with in normal tissue and was correlated with poor prognosis. M0 macrophages, M2 macrophages, resting mast, γ δ T and CD8+ T cells were the top five immune cells with the highest degrees of infiltration in osteosarcoma. In addition, BMPR2 expression level showed a significant negative correlation with the gene markers of CD8+ T cells, monocytes and M2 macrophages. Low levels of infiltrating CD8+ T cells, monocytes or M2 macrophages in osteosarcoma was significantly associated with poor survival. These data suggested that CD8+ T cells, monocytes and M2 macrophages play significant roles in the establishment of the immune microenvironment of osteosarcoma. High BMPR2 expression was associated with poor prognosis and low infiltration of CD8+ T cells, monocytes and M2 macrophages in osteosarcoma. Hence, BMPR2 can be considered a biomarker of the immune infiltration, metastasis and prognosis of osteosarcoma.     

Elevated expression of flotillin-1 is associated with lymph node metastasis and poor prognosis in early-stage cervical cancer

Accumulating evidence has revealed that the expression of the lipid raft protein flotillin-1 is elevated in various human cancers, but the role flotillin-1 plays in the carcinogenesis of cervical cancer remains unclear. The expression profile of flotillin-1 was assayed using real-time PCR, western blotting, and immunohistochemical (IHC) staining in cervical cancer cell lines and cancer tissues with paired adjacent noncancerous cervical tissues. The expression of flotillin-1 protein was detected by IHC staining in a large cohort of 308 paraffin-embedded cervical cancer tissues. Ectopic expression and the short hairpin RNA interference approach were employed to determine the role of flotillin-1 in cervical cancer cell metastasis and the possible mechanism involved. Flotillin-1 expression protein and mRNA were significantly upregulated in cervical cancer cell lines and cancer tissues; elevated expression of flotillin-1 protein in early-stage cervical cancer was significantly associated with pelvic lymph node metastasis (P < 0.001), and was an independent predictive factor of poor overall survival. Moreover, flotillin-1 up- and downregulation remarkably affected cervical cancer cell motility and invasion, respectively, through epithelial-mesenchymal transition (EMT) regulated by the Wnt/β-catenin and nuclear factor-κB (NF-κB) pathways. Our results suggest that flotillin-1 facilitates cervical cancer cell metastasis through Wnt/β-catenin and NF-κB pathway-regulated EMT and that the flotillin-1 expression profile serves not only as novel predictor of pelvic lymph node metastasis, but also as neoteric risk factor for patients with early-stage cervical cancer.     

Caveolin-1表达及其与胃癌生物学行为关系的研究

目的 :探讨候选抑癌基因Caveolin 1在胃癌中的表达及其与胃癌生物学行为的关系。方法 :应用免疫组织化学SABC法对 43例胃癌存档蜡块组织及 15例胃炎活检标本进行Caveolin 1蛋白检测 ,采用 χ2 检验进行统计学分析。结果 :Caveolin 1在胃癌的阳性表达率为 3 0 2 % ( 13 / 43 ) ,明显低于胃炎组 10 0 % ( 15 / 15 ) ,P <0 0 1;其中高分化腺癌、中分化腺癌、低分化腺癌中的阳性率分别为 6/ 6、3 / 5、17 4% ( 4 / 2 3 ) ,而印戒细胞癌及黏液腺癌中无阳性表达 ,五者之间差异有统计学意义 ,P <0 0 1;浸润至浆膜或超过浆膜组的Caveolin 1表达阳性率13 8% ( 4 / 2 9)明显低于未达浆膜组 64 3 %( 9/ 14 ) ,两组之间差异有统计学意义 ,P <0 0 1;淋巴结转移组Caveolin 1阳性率 7 7%( 1/ 13 )明显低于无淋巴结转移组 40 0 %( 12 / 3 0 ) ,两组之间差异有统计学意义 ,P <0 0 5。结论 :候选抑癌基因Caveolin 1的表达与胃癌细胞的分化、浸润和转移密切相关 ,对胃癌生物学行为评估有重要意义     

Pokemon mRNA在乳腺癌组织中的表达及意义

目的探讨乳腺癌组织中Pokemon mRNA的表达及其意义。方法采用原位杂交法,检测45例乳腺癌、20例癌旁乳腺组织、20例正常乳腺组织中Pokemon mRNA的表达,并对其与乳腺癌临床病理特征的关系进行分析。结果 Pokemon mRNA主要表达于乳腺癌细胞质。乳腺癌组织中Pokemon mRNA表达率为71.11%（32/45）,显著高于癌旁乳腺组织30.00%（6/20）和正常乳腺组织20.00%（4/20）,P〈0.05。有腋窝淋巴结转移组Pokemon mRNA表达率（90.00%）显著高于无腋窝淋巴结转移组（56.00%）,P〈0.05。组织学分级Ⅲ级组织Pokemon mRNA表达率（100.00%）显著高于Ⅰ级组织（53.33%）,P〈0.05。结论 Pokemon mRNA过度表达可能在乳腺癌发生中起重要作用,且与腋窝淋巴结转移及其组织学分级相关。     

赖氨酰氧化酶和基质金属蛋白酶2在胃癌组织中的表达及其与胃癌转移的关系

目的 探讨赖氨酰氧化酶(LOX)和基质金属蛋白酶2(MMP-2)在胃癌组织中的表达及其与胃癌转移的关系.方法 收集手术切除新鲜胃癌及相应的癌旁胃组织,用逆转录聚合酶链反应(RT-PCR)方法检测LOX mRNA和MMP-2 mRNA表达,Western blot方法检测LOX蛋白和MMP-2蛋白表达.结果 LOX mRNA在胃癌组织和癌旁胃组织中的相对表达量分别为0.5328±0.1367和0.2436±0.1372(P<0.01),MMP-2 mRNA在胃癌组织和癌旁胃组织中的相对表达量分别为0.7980±0.1571和0.3231±0.1672(P<0.05);LOX mRNA在有淋巴结转移和无淋巴结转移的胃癌组织中的相对表达量分别为0.6336±0.1547和0.4914±0.1408,MMP-2 mRNA在有淋巴结转移和无淋巴结转移的胃癌组织中的相对表达量分别为0.8762±0.1381和0.6362±0.1936,差异均有统计学意义(均P<0.05).LOX蛋白在胃癌组织和癌旁胃组织中的相对表达量分别为0.5237±0.1426和0.2972±0.1480(P<0.05),在有淋巴结转移和无淋巴结转移的胃癌组织中相对表达量分别为0.6880±0.1263和0.4944±0.1217(P<0.05);MMP-2蛋白在胃癌组织和癌旁胃组织中的相对表达量分别为0.7365±0.1607和0.3321±0.1480(P<0.05),在有淋巴结转移和无淋巴结转移的胃癌组织中相对表达量分别为0.8097±0.1632和0.5132土0.1366(P<0.05).胃癌组织中,LOX mRNA与MMP-2 mRNA、LOX蛋白与MMP-2蛋白相对表达量呈正相关(r=0.873,P<0.001;r=0.881,P<0.001);在伴有淋巴结转移的胃癌组织中,LOX mRNA与MMP-2 mRNA、LOX蛋白与MMP-2蛋白相对表达量也呈正相关(r=0.923,P<0.001;r=0.972,P<0.001).结论 LOX和MMP-2在胃癌组织中的表达明显高于相应的癌旁胃组织;在伴有淋巴结转移的胃癌组织中,LOX和MMP-2表达高于无淋巴结转移的胃癌组织,且LOX和MMP-2表达呈正相关,提示LOX和MMP-2对胃癌的发生和转移有促进作用,且两者可能有协同作用.

Abstract：

Objective To compare the expressions of lysyl oxidase (LOX) and matrix metalloproteinases-2 (MMP-2) in gastric cancer and pericancerous tissues, in gastric cancers.with and without lymph node metastasis, and to analyze the effects of LOX and MMP-2 on tumor invasion and metastasis.Methods Gastric cancer and pericancerous tissues were collected from 46 patients who underwent surgery.Levels of LOX and MMP-2 mRNA were detected by RT-PCR.Protein abundance of LOX and MMP-2 was examined using Western blot.Results Expressions of LOX and MMP-2 mRNA, and protein in 46 gastric cancers were significantly higher than that in 46 pericancerous tissues.In gastric cancer with lymph node metastasis, the levels of LOX and MMP-2 mRNA and protein were higher than those in gastric cancers without lymph node metastasis ( P < 0.05 ).In the groups of gastric cancer with lymph node metastasis, expression of LOX was positively correlated with MMP-2 protein expression ( P < 0.01 ).Conclusions Expressions of LOX and MMP-2 in gastric cancer tissues are significantly higher than that in pericancerous tissues.The expressions of LOX and MMP-2 in gastric cancer with lymph node metastasis are higher than that in gastric cancer without lymph node metastasis.Expressions of LOX and MMP-2 are positively correlated.The results suggest that LOX and MMP-2 may promote the growth and metastasis of gastric cancer.     

LINC00958 通过上调VEGF-C 表达促进宫颈癌细胞的恶性生物学行为及小鼠模型的淋巴结转移

目的：探讨LINC00958/血管内皮生长因子 C（VEGF-C）信号通路在宫颈癌的淋巴管生成和淋巴转移中的作用。方法：从2020 年9月至2022 年9月期间在河南省人民医院接受手术的患者中收集了42 例宫颈癌组织标本,通过qPCR检测宫颈癌组织和宫颈癌细胞（Hela、C33A、SiHa、Caski）中LINC00958 的表达情况。将LINC00958 过表达载体（LINC00958 组）或对照载体（CMV 组）转染Caski 细胞,敲减LINC00958（shLINC00958 组）、VEGF-C（shVEGF-C 组）的shRNA 序列或阴性对照shRNA （shNC 组）转染SiHa 细胞。分别通过CCK-8法、Transwell 实验检测过表达或敲减LINC00958 对宫颈癌细胞增殖、迁移和侵袭的影响。观察转染后细胞的培养上清液对人淋巴管内皮细胞（HLEC）淋巴管形成能力的影响。建立小鼠腘淋巴结转移模型,观察过表达LINC00958 或同时敲减VEGF-C对宫颈癌淋巴结转移的影响。结果：LINC00958 在宫颈癌组织中呈高表达（P<0.001）,高水平的LINC00958 与大肿瘤、晚期肿瘤分级、浸润深度和淋巴转移有关联（P<0.05 或P<0.01）。与正常人宫颈上皮细胞ende1617相比,宫颈癌细胞中LINC00958 水平均显著升高（P<0.01 或P<0.001）。shLINC00958 组SiHa 细胞的增殖、迁移、侵袭能力及其培养上清液的促HLEC淋巴管形成能力均显著低于shNC 组（P<0.05、P<0.01 或P<0.001）,LINC00958 组Caski 细胞的增殖、迁移、侵袭能力及其培养上清液的促HLEC淋巴管形成能力显著高于CMV组（P<0.05、P<0.01或P<0.001）。通过RNA下拉、RNA免疫沉淀实验发现宫颈癌细胞中LINC00958 能够特异性结合VEGF-C。LINC00958+shVEGF-C 组Caski 细胞的增殖、迁移、侵袭能力及其培养上清液的促淋巴管形成能力显著低于LINC00958 组（P<0.01 或P<0.001）;在小鼠腘淋巴结转移模型中,LINC00958+ shVEGF-C 组中小鼠腘窝淋巴结的体积和VEGF-C 蛋白、N-cadherin 蛋白以及LYVE-1的阳性细胞比例均显著低于LINC00958 组（均P<0.001）。结论：LINC00958通过直接与VEGF-C蛋白相互作用增强宫颈癌细胞的增殖、侵袭、淋巴管生成能力,促进小鼠腘淋巴结转移模型的淋巴结转移。     

Cycloxygenase-2 inhibition augments the efficacy of a cancer vaccine.

Tumor-derived cyclooxygenase-2 (COX-2) and its product, prostaglandin E2, exert strong immunoinhibitory effects that block dendritic cell function and CD4+ and CD8+ T-cell proliferation and function. We have shown previously that the addition of an oral COX-2 inhibitor to immunogene therapy using IFN-beta markedly augmented therapeutic efficacy in murine tumor models. In this study, we hypothesized that COX-2 inhibition might also augment an antitumor vaccination strategy. Mice bearing tumors derived from TC1 cells, a tumor line that expresses the human papillomavirus (HPV) E7 protein, were thus vaccinated with an adenoviral vector expressing HPV E7 protein (Ad.E7). This vaccine approach effectively generated E7-specific CD8+ cells and slowed the growth of small tumors but had little effect on large tumors. However, feeding mice with the COX-2 inhibitor, rofecoxib, restored the effectiveness of the vaccine against large tumors and prolonged survival. This effect was accompanied by a larger percentage of E7-specific CD8+ cells in the regional draining lymph nodes and a markedly increased number of tumor-infiltrating E7-specific CD8+ cells (as determined by flow cytometry) and total CD8+ T cells (as determined by immunohistochemical staining). Increased immunocyte trafficking was likely mediated by the generation of a Th1-type tumor microenvironment because COX-2 inhibition increased expression levels of mRNA for IFN-gamma, interleukin-12, IP-10, and MIG while lowering the expression of vascular endothelial growth factor within tumors. This study shows that the effectiveness of a cancer vaccine can be significantly improved by adding COX-2 inhibition.