目的 分析甲状腺乳头状癌(PTC)超声图像特征及BRAF V600E突变与颈部淋巴结转移的关系.方法 选取于我院行甲状腺癌根治术患者316例,根据甲状腺癌术后病理分期(pTNM)结果,将pN0(无颈部淋巴结转移)者归为未转移组(160例),pN1(有颈部淋巴结转移)者归为转移组(156例),比较两组超声图像特征、临床资料、BRAF V600E突变情况的差异.采用多因素Logistic回归分析PTC超声图像特征、BRAF V600E突变及各临床资料与颈部淋巴结转移的关系.结果 转移组98例(62.8％)发生BRAF V600E突变,未转移组121例(75.6％)发生BRAF V600E突变,两组BRAF V600E突变占比比较,差异有统计学意义(P<0.05).转移组与未转移组年龄及病灶数目、大小、沙粒样钙化、纵横比、边缘、与被膜关系、累及腺叶比较,差异均有统计学意义(均P<0.05).多因素Logistic回归分析显示,病灶大小、纵横比、与被膜关系、累及腺叶均为颈部淋巴结转移的危险因素(OR=3.606、4.061、2.149、8.578,均P<0.05),年龄、沙粒样钙化、病灶边缘均为颈部淋巴结转移的保护因素(OR=0.364、0.389、0.409,均P<0.05).结论 PTC颈部淋巴结转移与年龄及病灶大小、沙粒样钙化、纵横比、边缘、与被膜关系、累及腺叶均有关,与性别、病灶数目、BRAF V600E突变无关.

目的 探讨超声引导下细针抽吸细胞学检查(FNAC)联合BRAFV600E基因检测对甲状腺微小乳头状癌(PTMC)的术前诊断价值.方法 选取我院经手术病理确诊的PTMC患者126例,共计126个结节,按结节二维超声所测最大径分为≤5 mm组31个,5～10 mm组95个,比较两组FNAC、BRAFV600E基因检测诊断结果,分析二者联合诊断价值.结果 5～10 mm组FNAC的诊断准确率(86.32％)明显高于≤5 mm组(58.06％),两组比较差异有统计学意义(P<0.05);5～10 mm组与≤5 mm组BRAFV600E基因突变率分别为92.63％、90.32％,两组比较差异无统计学意义;FNAC联合BRAFV600E基因检测诊断≤5 mm组、5～10 mm组的准确率分别为93.54％、97.78％,均明显高于单独FNAC诊断,差异均有统计学意义(均P<0.05).结论 FNAC联合BRAFV600E基因检测可提高PTMC术前诊断准确率,且对5～10 mm PTMC诊断准确率更高.

Objective To investigate the effect of transmembrane protein 196 (TMEM196) on the migration and invasion of breast cancer cells. Methods The Cancer Genome Atlas (TCGA) and Oncomine database were used to analyze the mRNA expression of TMEM196 in breast cancer. The expression of TMEM196 in breast cancer cells was detected by quantitative real time-polymerase chain reaction (RT-qPCR). Kaplan-Meier database was employed to analyze the effect of TMEM196 on the prognosis of breast cancer patients. The protein level and prognostic effect of TMEM196 were analyzed by tissue microarray. The cohort included 70 paracancerous samples and 160 invasive breast cancer samples from the patients (53.61±13.13 years old) undergoing surgical resection from January 2001 to August 2004, with a follow-up period of 2 to 150 months. Cell proliferation experiment, scratch healing test and Transwell assay were performed to determine the effect of TMEM196 on the proliferation, migration and invasion of breast cancer cells. Western blot assay was used to explore the regulation of TMEM196 on the expression of β-catenin and matrix metalloproteinases 7 (MMP7). Results The mRNA and protein levels of TMEM196 were significantly decreased in breast cancer (P < 0.05), and notably, this phenomenon was most obvious in triple negative breast cancer (TNBC) cell line MDA-MB-231 and TNBC tissues (P < 0.05). Lower expression of TMEM196 was associated with poor overall survival (OS) (HR=0.524, 95%CI: 0.293~0.936, P=0.040). In addition, overexpression of TMEM196 could significantly inhibit the proliferation, migration and invasion of breast cancer cells(P < 0.05), while its knockdown can significantly enhances the above abilities(P < 0.05). The results of Western blot assay indicated that TMEM196 inhibited the β-catenin/MMP7 signaling pathway and reduced the motility of breast cancer cells(P < 0.05). On the contrary, the tumor-promoting effect which caused by knockdown of TMEM196 could be eliminated after adding β-catenin inhibitor(P < 0.05). Conclusion TMEM196 inhibits the migration and invasion of breast cancer cells by regulating the β-catenin/MMP7 signaling pathway.

Objective:To analyze the immune implication of NR3C2 gene in breast cancer using bioinformatics methods and construct a prognostic model.Methods:(1) The breast cancer cohort in the Cancer Genome Atlas (TCGA) database and the GSE42568 cohort in the Gene Expression Omnibus (GEO) database were used as training set (113 paracancerous samples and 1 019 breast cancer samples) and testing set (17 paracancerous samples and 104 breast cancer samples), respectively. The mRNA level of NR3C2 was compared between adjacent tissue and tumor tissue samples in the above 2 cohorts. The effect of NR3C2 expression on recurrence-free survival (RFS) was analyzed in cohorts of TCGA and Kaplan-Meier plotter(4 929 breast cancer samples), respectively. (2) The Gene Set Enrichment Analysis (GSEA) was employed to explore the potential biological functions of NR3C2. The 24 immune cells were quantitively accessed by single sample gene set enrichment analysis (ssGSEA) and the correlation of NR3C2 with 24 immune cells and 70 immunomodulator genes were conducted by Pearson coefficient. (3) A prognostic model was constructed by NR3C2-related immunomodulator genes in the TCGA cohort through multivariate stepwise Cox regression. The TCGA cohort was divided into two groups (high-risk group and low-risk group) by median risk value and RFS was compared between two groups. The sensitivity and specificity of the model was calculated using receiver operating characteristic (ROC) curves and validated in the GSE42568 cohort. Combined with other clinical parameters, the independent prognostic performance of this model was analyzed by multivariate Cox regression. (4) A nomogram was constructed based on pathological stage and risk value in TCGA cohort. The calibration curve was used to evaluate its accuracy and the predictive accuracy of different parameters was compared by time-dependent area under curve (tAUC). (5) In order to verify whether the NR3C2 mRNA expression is consistent with NR3C2 protein expression, we collected clinical specimens from three breast cancer patients who underwent surgical resection in the Department of Breast and Thyroid Surgery of the Army Medical Center in September 2021. The protein expression of NR3C2 in the paracancerous tissue and cancer tissue was detected by Western blot analysis.Results:(1) In TCGA cohort, the mRNA expression of NR3C2 in breast cancer tissue was significantly lower than that in paracancerous tissues (2.59±0.43 vs 0.98±0.62, t=35.990, P<0.001). In GSE42568 cohort, the mRNA expression of NR3C2 in breast cancer tissue was significantly lower than that in paracancerous tissues (5.35±1.47 vs 3.32±1.12, t=7.096, P<0.001). The results of survival analysis showed that in TCGA cohort and Kaplan-Meier plotter cohort, NR3C2 expression was positively correlated with RFS in breast cancer patients (HR=0.667, 0.725; 95%CI: 0.458-0.972, 0.653-0.804; both P<0.050). (2) GSEA results suggested that NR3C2 was mostly involved in JAK-STAT and TGF-β signaling pathways related to immunity. Correlation analysis found that the mRNA expression of NR3C2 was significantly correlated with 19 immune cells and 43 immunomodulator genes (all P<0.050). (3) The 43 NR3C2-related immunomodulator genes were included in Cox regression to construct a prognostic model which composed of 13 immunomodulator genes with a risk cutoff value equal to 0.988. Survival analysis showed that in TCGA cohort and GSE42568 cohort, the RFS in high-risk group was significantly lower than that in low-risk group (HR=2.682, 2.389; 95%CI: 1.839-3.910, 1.343-4.248; both P<0.010). ROC indicated that the AUC was 0.758 and 0.618 in TCGA cohort and GSE42568 cohort, respectively (95%CI: 0.662-0.857, 0.545-0.758; sensitivity: 0.833, 0.538; specificity: 0.614, 0.714; both P<0.010). Multivariate Cox regression showed that the risk score of this model could serve as an independent prognostic factor for breast cancer in TCGA cohort and GSE42568 cohort (HR=1.259, 1.163; 95%CI: 1.187-1.336, 1.068-1.266; both P<0.001). (4) The nomogram constructed based on the pathological stage and risk value could predict the 3-year, 5-year and 8-year RFS of breast cancer patients. The calibration curve suggested that it has good predictive accuracy and tAUC indicated that it is superior to pathological stage and a prognostic model. (5) Western blot analysis showed that the protein expression of NR3C2 was significantly decreased in breast cancer tissues.Conclusion:NR3C2 is a potential immunotherapeutic target and prognostic biomarker in breast cancer patients.