Spatial transcriptome analysis of long non-coding RNAs reveals tissue specificity and functional roles in cancer / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Long non-coding RNAs (lncRNAs) play a significant role in maintaining tissue morphology and functions, and their precise regulatory effectiveness is closely related to expression patterns. However, the spatial expression patterns of lncRNAs in humans are poorly characterized. Here, we constructed five comprehensive transcriptomic atlases of human lncRNAs covering thousands of major tissue samples in normal and disease states. The lncRNA transcriptomes exhibited high consistency within the same tissues across resources, and even higher complexity in specialized tissues. Tissue-elevated (TE) lncRNAs were identified in each resource and robust TE lncRNAs were refined by integrative analysis. We detected 1 to 4684 robust TE lncRNAs across tissues; the highest number was in testis tissue, followed by brain tissue. Functional analyses of TE lncRNAs indicated important roles in corresponding tissue-related pathways. Moreover, we found that the expression features of robust TE lncRNAs made them be effective biomarkers to distinguish tissues; TE lncRNAs also tended to be associated with cancer, and exhibited differential expression or were correlated with patient survival. In summary, spatial classification of lncRNAs is the starting point for elucidating the function of lncRNAs in both maintenance of tissue morphology and progress of tissue-constricted diseases.

Correlations of expressions of tissue inhibitor of matrix metalloproteinase-1 and fibronectin 1 in pregnancy associated breast cancer with expression of E-cadherin and prognosis / 肿瘤研究与临床

Objective:To investigate the expressions of tissue inhibitor of matrix metalloproteinase-1 (TIMP1) and fibronectin 1 (FN1) in pregnancy associated breast cancer (PABC) and their correlations with expression of E-cadherin (E-cad).Methods:The clinicopathological data of 55 PABC patients in Binzhou People's Hospital Affiliated to Shandong First Medical University from January 2011 to December 2020 were retrospectively analyzed. Immunohistochemistry was used to detect expressions of TIMP1, FN1 and E-cad in cancer tissues and corresponding paracancerous tissues (>3 cm from the edge of the tumor foci). The expressions of TIMP1 and FN1 proteins in fresh intraoperative frozen cancer tissues and paracancerous tissues of 10 PABC patients were detected by Western blotting. The correlations of TIMP1 and FN1 expressions with clinicopathological characteristics of patients were analyzed by χ2 test, the correlation of TIMP1 and FN1 expressions with E-cad expression was analyzed by Spearman method, and the correlation of TIMP1 and FN1 expressions with survival was analyzed by Kaplan-Meier method. Results:The positive rates of TIMP1 and FN1 in PABC tissues were 72.7% (40/55) and 58.2% (32/55), and 25.5% (14/55) and 18.2% (10/55) in paracancerous tissues, and the differences were statistically significant ( χ2 values were 24.59 and 18.64, both P < 0.001). The results of Western blotting showed that the relative expressions of TIMP1 and FN1 proteins in the fresh cancer tissues of 10 PABC patients was higher than those in the corresponding paracancerous tissues (1.60±0.76 vs. 0.62±0.29, 1.31±0.62 vs. 0.44±0.15), and the differences were statistically significant ( t values were 5.92 and 4.86, both P < 0.001). The expressions of TIMP1 and FN1 in PABC tissues were correlated with estrogen receptor expression, Ki-67 positivity index, TNM stage and lymph node metastasis (all P < 0.05). The expressions of TIMP1 and FN1 were negatively correlated with expression of E-cad in PABC ( r values were -0.471 and -0.432, both P < 0.001). Five cases were lost to follow-up, and the remaining 50 cases had a median follow-up time of 43 months (12-90 months). Among the 50 cases, 36 cases were TMP1-positive and 29 cases were FN1-positive. The overall survival of TIMP1-negative group and FN1-negative group were better than those of the corresponding positive group ( χ2 values were 4.49 and 6.06, both P < 0.05); the median overall survival time of TIMP1-positive group and FN1-positive group were 51 months (95% CI 37-65 months) and 43 months (95% CI 32-53 months), while that of TIMP1-negative group and FN1-negative group were 89 months (95% CI 84-93 months) and 87 months (95% CI 85-92 months). Conclusions:TIMP1 and FN1 expressions are elevated in PABC tissues and negatively correlated with E-cad expression, TIMP1 and FN1 may be involved in PABC invasion through epithelial-mesenchymal transition and affect the prognosis of patients.

Detection of epidermal growth factor receptor mutations in non-small cell lung cancer by immunohistochemistry / 中南大学学报(医学版)

OBJECTIVES@#To evaluate the sensitivity and specificity of immunohistochemistry (IHC) for detecting common epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) and to estimate the cost-effectiveness of IHC testing.@*METHODS@#A total of 208 NSCLC patients were included in the trial, and the EGFR mutation status in the patients were detected by PCR and IHC. Two mutation-specific antibodies against the most common exon 19 deletion (clone SP111) and exon 21 L858R mutation (clone SP125) were tested by using automated immunostainer. A cost-effectiveness analysis model was built for the analysis of optimal detection scheme.@*RESULTS@#With a cutoff value of IHC 1+, the overall sensitivity and specificity of the IHC-based method compared with the PCR-based method were 81.7% (95% CI 72.4% to 89.0%) and 94.7% (95% CI 92.6% to 99.5%), respectively. EGFR 19del mutation was detected by SP111 antibody with a sensitivity of 65.9% (95% CI 49.4% to 79.9%) and specificity of 98.8% (95% CI 95.7% to 99.9%). EGFR L858R mutation was detected by SP125 antibody with a sensitivity of 94.2% (95% CI 84.1% to 98.8%) and specificity of 99.4% (95% CI 96.5% to 100%). The IHC and PCR cost ratio needed to be 1-to-3 or more in our patients to economically justify before the use of IHC.@*CONCLUSIONS@#The study confirms an excellent specificity with fairly good sensitivity of IHC and mutation-specific antibodies for common EGFR mutations. It is cost-effective to use IHC method to detect EGFR mutation first when the IHC and PCR cost ratio is 1-to-3 or more in Chinese populations.

Invasive breast lobular carcinoma with extracellular mucin: a clinicopathological analysis / 中华病理学杂志

Objective@#To study the clinicopathological features of invasive lobular carcinoma (ILC) of the breast with extracellular mucin and outcomes of patients.@*Method@#Clinicopathological features and clinical follow-up (39-123 months and a median follow-up of 55 months) of seven ILC with extracellular mucin were obtained. Hematoxylin-and-eosin (H&E) and immunohistochemistry (IHC) stained sections were reviewed, and fluorescence in situ hybridization (FISH) assay was performed for tumors with HER2 IHC 2+. Patient prognosis was analyzed and literatures related to ILC with extracellular mucin were reviewed.@*Results@#All seven patients were female, aged from 43 to 73 years (median age, 55 years). The tumors ranged in size from 1 to 5 cm (median size 2 cm). All seven cases were of histological grade 2. Most areas of the tumors presented with the morphology of classic ILC, and variable amount of extracellular mucin were observed focally. In six cases, part of the tumor cells contained intracellular mucin, and the nucleus were pushed to one side of the cells, creating the impression of signet-ring cells. Two patients had lymph node metastases at diagnosis, and developed liver and bone metastases at 38th and 48th month, respectively, after surgery, and died at 48th and 123th month, respectively. While the other five patients, except one lost to follow-up, had been disease-free during the follow-up period. IHC results showed estrogen receptor (ER) and progesterone receptor (PR) positivity in 7/7 and 6/7 cases, respectively. Tumors of six patients were HER2 IHC 0/1+. The remaining one was HER2 IHC 2+, while FISH assay revealed HER2 gene amplification in that tumor. The proportion of cases with HER2-positivity was 1/7. The proliferation index Ki-67 ranged from less than 5% to 30%, and Ki-67 less than or equal to 10% were in 5/7 cases. According to the 2013 St. Gallen International Expert Consensus on breast cancer, all tumors were of luminal types; of those, two were luminal A and five were luminal B.@*Conclusions@#ILC with extracellular mucin tends to occur in women over 50 years old. All tumors in the study are grade 2 classic ILC, with signet-ring cells as a common feature. All seven tumors are classified as luminal types, with luminal B as the main molecular subtype.

Expression of epithelial cell adhesion molecule and molecular subtypes and prognosis of breast cancer / 中南大学学报(医学版)

OBJECTIVE@#To investigate clinicopathological and prognostic significance of epithelial cell adhesion molecule (EpCAM) expression in breast cancer and its molecular subtypes.
@*METHODS@#The expression of EpCAM in 835 patients with breast invasive ductal carcinoma was detected by immunohistochemical Max VisionTM method, and its correlation with clinical pathological features and prognosis was analyzed.
@*RESULTS@#The positive expression of EpCAM was related to histological grade, lymph node metastasis, tumor size, clinical stage, the expression of estrogen receptor (ER), progesterone receptor (PR) and HER2 (P<0.05). The positive expression rates of EpCAM were 19.2%, 73%, 48.9%, 72.2%, and 62.1%, in Luminal A, Luminal B (HER2-), Luminal B(HER2+), HER2+, and triple-negative subtype, respectively. Log-rank test and univariate COX analysis showed that the EpCAM expression was associated with a poor prognosis in all patients (P<0.001), as well as the triple-negative subtype, luminal B subtype (HER2-), and HER2+ subtype (P<0.05). Multivariate COX analysis showed that EpCAM expression was associated with the survival in patients with the triple-negative or HER2+ subtype (P<0.05).
@*CONCLUSION@#EpCAM may be associated with progress of breast cancer. It is an independent prognostic factor in breast cancer, especially in the triple-negative and HER2+ subtypes.

Expressions of epithelial cell adhesion molecule, vimentin and N-cadherin in molecular subtypes of breast cancer and the correlation among them / 中南大学学报(医学版)

To examine the expressions of epithelial cell adhesion molecule (EpCAM), vimentin and N-cadherin in breast cancer and its molecular subtypes, and to explore the correlation among them.
 Methods: The expressions of EpCAM, vimentin and N-cadherin were detected by immunohistochemistry in 835 patients with breast cancer, and their correlations with clinical pathological features and prognosis were analyzed.
 Results: The expression rates of EpCAM, vimentin and N-cadherin in the patients were 53.4%, 11.4% and 9.7% respectively, which were increased with the increase in tumor size, histological grade, lymph node size, tumor node stage of metastases classification, estrogen receptor and progesterone receptor levels (all P<0.05). The positive expression rates for EpCAM protein in luminal A, luminal B (HER2-), luminal B (HER2+), HER2 overexpression and triple-negative subtypes were 19.2%, 73.0%, 48.9%, 72.2%, and 62.1% respectively; for vimentin were 3.9%, 11.4%, 14.1%, 11.1%, and 20.5% respectively; for N-cadherin were 7.0%, 5.7%, 12.0%, 12.2% and 17.4% respectively, with statistical difference (all P<0.05). EpCAM expression was positively correlated with vimentin and N-cadherin in patients with breast cancer and triple-negative breast cancer.
 Conclusion: EpCAM is overexpressed in triple-negative subtype of breast cancer and it is associated with epithelial-mesenchymal transition markers, which might be related to breast cancer progression and metastasis.

Clinicopathological significance of combined immunohistochemical analysis of PTEN, p53 and EGFR in breast carcinoma / 临床与实验病理学杂志

Purpose To study clinical pathological significance of the expression of PTEN, p53 and epidermal growth factor receptor ( EGFR) in breast carcinoma and their correlation. Methods Immunohistochemical MaxVision method was used to detect the expres-sion of PTEN, p53, EGFR in 209 cases of infiltrating ductal carcinoma of breast and 40 cases of benign breast diseases. Results The over-expression rate of PTEN, p53 and EGFR protein in breast carcinoma was 57. 9%, 55. 0% and 38. 3% respectively, which was significantly different from those in benign breast diseases (P<0. 05). The expression of PTEN, p53 and EGFR in breast cancer was correlated with tumor size, histological grade, lymph node metastasis, ER status, PR status and molecular subtype (P<0. 05). There was an association between PTEN or p53 and TNM stage, PTEN or EGFR and HER-2 status. There was a negative correlation in the protein expression of PTEN vs EGFR or PTEN vs p53 (P<0. 01). There was a positive correlation between EGFR and p53 protein ex-pression (P=0. 000). The expression of PTEN or EGFR was correlated with the prognosis of breast cancer, but not independent prog-nostic factors. The survival rate of patients with PTEN- /p53 + /EGFR+ was lower than those with other combined expression of PTEN/p53/EGFR (P=0. 008). Conclusions Low or loss expression of PTEN, p53 mutation and EGFR over-expression may be coordinate-ly involved in the pathogenesis and progression of breast cancer. The combined detection of these markers may play an important role in making treatment and indicating prognosis for breast cancer patients.

Expression of PTEN, p53 and EGFR in the molecular subtypes of breast carcinoma and the correlation among them / 中南大学学报(医学版)

OBJECTIVE@#To study the expression of PTEN, p53 and epidermal growth factor receptor (EGFR) in the molecular subtypes of breast carcinoma and to evaluate the correlations with triple-negative breast cancer.
@*METHODS@#Immunohistochemical MaxVision(TM) method was used to detect the expression of PTEN, p53, EGFR, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) in 291 cases of infiltrating ductal carcinoma of breast. 
@*RESULTS@#The positive expression of PTEN, p53 and EGFR protein in breast carcinoma was 57.0%, 57.0% and 38.5%, respectively, which were significantly different from those in benign breast diseases (P<0.05). The expression of PTEN or EGFR in breast cancer was correlated with tumor size, histological grade, lymph node metastasis, TNM stage, ER and HER2 status (P<0.05); the expression of p53 was correlated with tumor size, histological grade and ER status (P<0.05). The difference of positive expression rates of PTEN, p53 and EGFR protein among different subtypes including luminal A, luminal B (HER2-), luminal B (HER2+), HER2 over-expression and triple-negative was statistically significant (P<0.05). There were close correlations among PTEN, p53 and EGFR in the triple-negative subtype (P<0.05).
@*CONCLUSION@#Low expression of PTEN and high expression of EGFR and p53 are observed in triple-negative breast cancer, which may synergistically contribute to the pathogenesis of triple-negative breast cancer.

Expression of estrogen receptor and progesterone receptor in intrahepatic and extrahepatic cholangiocarcinoma / 肿瘤研究与临床

ObjectiveTo investigate the expression of estrogen receptor(ER)and progesterone receptor(PR)in intrahepatic cholangiocarcinoma(ICCA)and extrahepatic cholangiocarcinoma(ECCA)comparatively.MethodsThe expression of ER and PR was detected in 24 specimens of ICCA and 34specimens of ECCA by immunohistochemistry assay.ResultsThe positive rates for ER and PR expression were 29 ％(7/24)and 46 ％(11/24)in ICCA specimens,respectively.3 cases among 9 cases of high histological grade,3 cases among 11 cases of middle histological grade and 1 case among 4 cases of low histological grade expressed ER positively,while 2 cases,7 cases and 2 cases expressed PR positively,respectively.The expression of ER and PR expressions showed no statistically significant differences between different histological grades of ICCA.ER and PR were all negative in 34 specimens of ECCA.Conclusion ER and PR show significant different expression between ICCA and ECCA,with exclusive positive expression in ICCA.ER and PR might be involved in the pathogeuesis of ICCA.