Objective To investigate the expression of insulin-like growth factor 2 mRNA binding protein 1(IGF2BP1)mRNA and patrilineal expression of genetic imprinting gene 10(PEG10)mRNA in endometrial carcinoma(EC)tissues and their correlation with proliferation gene expression and prognosis.Methods A total of 100 EC patients diagnosed and treated in Hubei Institute of Technology Affiliated Maternal and Child Health Hospital from January 2017 to January 2019 were selected.The expression of IGF2BP1 mRNA,PEG 10 mRNA,proliferating cell nuclear antigen(PCNA),cyclin D1,and cyclin dependent kinase 4(CDK4)mRNA were examined by real-time fluorescence quantitative PCR.The expressions of IGF2BP1 and PEG 10 in tissues were detected by immunochemistry,and their correlation were analyzed by Pearson correlation analysis.Kaplan-Meier curve analysis were used to analyze the prognostic differences in EC patients with different IGF2BP1 and PEG 10 expression groups.COX regression was used to analyze the prognostic factors of EC patients.Results The expression levels of IGF2BP1 mRNA(1.84±0.33),PEG10 mRNA(2.12±0.40),PCNA mRNA(3.14±0.42),cyclinD1 mRNA(2.81±0.36)and CDK4 mRNA(2.37±0.34)in EC cancer tissues were higher than those in adjacent tissues(0.78±0.21,0.91±0.25,0.74±0.13,0.67±0.21,0.59±0.18),and the differences were significant(t=25.652～54.588,all P＜0.05).The positive rates of IGF2BP1(70.00％)and PEG 10(72.00％)in cancer tissues were higher than those in adjacent tissues(10.00％,9.00％),and the differences were statistically significant(x2=75.000,82.363,all P＜0.05).The expressions of IGF2BP1 mRNA and PEG10 mRNA in EC tissues were positively correlated with PCNA mRNA,cyclinD1 mRNA,and CDK4 mRNA(r=0.562～0.625,all P＜0.05).There was a significant positive correlation between IGF2BP1 mRNA and PEG10 mRNA expression in EC tissues(r=0.663,P＜0.05).The positive rates of IGF2BP1(86.49％,87.50％)and PEG10(89.19％,90.63％)protein expression in EC cancer tissues with FIGO stage Ⅲ and combined lymph node metastasis were higher than those in FIGO stage Ⅰ-Ⅱ(60.321％,61.90％)and without lymph node metastasis(61.77％,63.24％),and the differences were sttistically significant(x2=6.863～8.608,all P＜0.05).The overall 3-year survival rate of IGF2BP1 positive patients[70.00％(49/70)]was lower than that of IGF2BP1 negative patients[90.00％(27/30)],the overall 3-year survival rate of patients in PEG10 positive patients[69.40(50/72)]was lower than that in PEG10 negative patients[92.86％(26/28)],and the differences were statistically significant(Log rankx2=4.133,5.491,P=0.042,0.019).FIGO stage Ⅲ(OR=1.449,95％CI:1.148～1.830),combined lymph node metastasis(OR=1.442,95％CI:1.124～1.850),IGF2BP1 positive(OR=1.637,95％CI:1.239～2.163),and PEG10 positive(OR=1.576,95％CI:1.136～1.187)were independent risk factors affecting the survival and prognosis of EC patients(all P＜0.05).Conclusion The expressions of IGF2BP1 and PEG 10 in EC were increased,and they were positively correlated with the proliferation gene expression.They may be potential tumor markers for prognostic evaluation of EC.

Multi-gene assays have emerged as crucial tools for risk stratification in early-stage breast cancer. This study aimed to evaluate the prognostic significance of the 21-gene recurrence score (RS) in Chinese patients with pN0-1, estrogen receptor-positive (ER
Biomarkers, Tumor/genetics* ; Breast Neoplasms/pathology* ; Female ; Humans ; Neoplasm Recurrence, Local/pathology* ; Neoplasm Staging ; Prognosis ; Receptor, ErbB-2/genetics* ; Receptors, Estrogen

Objective:To investigate the expression of microRNA (miR) -26a in human skin fibroblasts during photoaging induced by ultraviolet A (UVA) , and to evaluate the effect of up-or down-regulation of miR-26a expression on the methylation level of the whole genome, the target gene enhancer of zeste homolog 2 (EZH2) and cell aging.Methods:Some human skin fibroblasts were irradiated with 10 J/cm 2 UVA once a day for 7 consecutive days, RNA was extracted on days 0, 3 and 7, and real-time quantitative reverse PCR (RT-PCR) was performed to determine the expression of miR-26a; miR-26a mimics and inhibitors were transfected into fibroblasts to up-or down-regulate the expression of miR-26a respectively, and fluorescence microscopy and RT-PCR were performed to determine the expression of miR-26a and evaluate the transfection efficiency. Some human skin fibroblasts were divided into 6 groups: blank control group receiving no treatment, UVA group treated with UVA irradiation according to the above method, miR-26a mimic group transfected with miR-26a-mimics, UVA+miR-26a mimic group transfected with miR-26a-mimics followed by UVA irradiation, miR-26a inhibitor group transfected with miR-26a inhibitors, UVA+miR-26a inhibitor group transfected with miR-26a inhibitors followed by UVA irradiation. On day 7, cells in each group were collected after the end of UVA irradiation. Then, flow cytometry was performed to detect cell cycle, DNA methylation quantitative detection kit was used to detect the methylation level of whole genome, RT-PCR was conducted to determine the mRNA expression of EZH2 (a histone-lysine N-methyltransferase enzyme) , DNA methyltransferase 1 (DNMT1) and miR-26a, and Western blot analysis was performed to determine the protein expression of EZH2 and DNMT1. Statistical analysis was carried out by using one-way analysis of variance and least significant difference- t test. Results:Compared with the unirradiated control group, the expression of miR-26a gradually increased in the UVA irradiation group over time during the culture, and there was a significant difference in the expression of miR-26a between the two groups after 7 days of UVA irradiation ( t=5.295, P < 0.05) . Strong fluorescence signals were observed in the miR-26a mimic-or miR-26a inhibitor-transfected fibroblasts, suggesting a high transfection efficiency. Flow cytometry showed that the proportion of cells at G1 phase significantly differed among the blank control group, UVA group, miR-26a mimic group, UVA+miR-26a mimic group, miR-26a inhibitor group, and UVA+miR-26a inhibitor group (52.82% ± 2.56%, 78.56% ± 4.34%, 53.63% ± 3.13%, 89.52% ± 4.17%, 54.39% ± 3.86%, 65.34% ± 4.78%, respectively; F=46.728, P < 0.01) , and significantly higher in the UVA group than in the blank control group ( t=8.848, P < 0.01) , higher in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group ( t=11.922, 3.154, P < 0.01, < 0.05, respectively) , and higher in the UVA+miR-26a inhibitor group than in the miR-26a-inhibitor group ( t=3.087, P < 0.05) , but significantly lower in the UVA+miR-26a inhibitor group than in the UVA group ( t=3.547, P < 0.05) . Detection of the genome-wide methylation level showed that the methylation level ( A450 value) significantly differed among the above groups (0.676 ± 0.024, 0.323 ± 0.043, 0.506 ± 0.035, 0.169 ± 0.024, 0.602 ± 0.036, 0.422 ± 0.029, respectively, F=97.402, P < 0.01) , and significantly lower in the UVA group than in the blank control group ( P < 0.01) , lower in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group (both P < 0.01) , and lower in the UVA+miR-26a inhibitor group than in the miR-26a inhibitor group ( P < 0.01) , but significantly higher in the UVA+miR-26a inhibitor group than in the UVA group ( P < 0.05) . RT-PCR and Western blot analysis showed significant differences in the mRNA and protein expression of EZH2 and DNMT1 respectively among the 6 groups (both P < 0.05) , which were significantly lower in the UVA group than in the blank control group ( P < 0.05) , lower in the UVA+miR-26a mimic group than in the miR-26a mimic group and UVA group (both P < 0.05) , and lower in the UVA+miR-26a inhibitor group than in the miR-26a inhibitor group ( P < 0.05) , but significantly higher in the UVA+miR-26a inhibitor group than in the UVA group ( P < 0.05) . Conclusion:In the UVA irradiation-induced photoaging of skin fibroblasts, miR-26a expression was up-regulated, cellular proliferative activity and genome-wide methylation level decreased; up-regulation of miR-26a expression could down-regulate the expression of its target gene EZH2 and methylation-related gene DNM1, and promote cell photoaging, while down-regulation of miR-26a expression could up-regulate the expression of EZH2 and DNMT1, and inhibit cell photoaging.

Purpose: This retrospective study aimed to evaluate the distribution pattern and prognostic value of 21-gene recurrence score (RS) in Chinese patients with mucinous breast cancer (MC) and compared with infiltrating ductal carcinoma (IDC). Materials and Methods: Patients diagnosed with MC or IDC from January 2010 to January 2017 were retrospectively recruited. Reverse transcriptase–polymerase chain reaction assay of 21 genes was conducted to calculate the RS. Univariate and multivariate analyses were performed to assess the association between RS and clinicopathological factors. Survival outcomes including disease-free survival (DFS) and overall survival (OS) were estimated by Kaplan-Meier method and compared by log-rank test. Results: The MC cohort included 128 patients and the IDC cohort included 707 patients. The proportions of patients with a low (RS < 18), intermediate (18-30), or high risk (RS > 30) were 32.0%, 48.4%, and 19.5% in MC cohort, and 26.9%, 46.8% and 26.3% in IDC cohort. The distribution of RS varied significantly according to different Ki-67 index and molecular subtype in both cohorts. Moreover, the receipt of chemotherapy was associated with RS in both cohorts. Among patients with MC, tumor stage was related to the DFS (p=0.040). No significant differences in DFS and OS were found among MC patients in different RS risk groups (OS, p=0.695; DFS, p=0.926). Conclusion RS was significantly related to Ki-67 index and molecular subtypes in MC patients, which is similar in IDC patients. However, RS was not able to predict DFS and OS in patients with MC.

Objective: The current study aimed to evaluate the predictive performances of anatomic staging system (AS) and prognostic staging system (PS) proposed in the 8th edition American Joint Committee on Cancer (AJCC) staging manual in patients with pure mucinous breast cancer (PMBC). Methods: Clinicopathologic features and follow-up information were collected from a total of 3628 patients with PMBC. Breast cancer-specific survival (BCSS) were compared among patients in different stage groups. Likelihood ratio (LR) χ², Akaike information criterion (AIC) and Harrell′s concordance index (C-index) were used to evaluate the predictive performances of AS and PS in PMBC. Results: In PMBC, BCSS was associated with tumor size (P=0.002), lymph node status (P=0.002), grade(P=0.003), PR status(P=0.017)and the receipt of radiation. Compared to AS, 1326 patients (37.54%) underwent stage change after applying PS, with 6.50% upstaged and 37.04% downstaged. There were significant differences in BCSS among patients of different stages under the AS and PS (P<0.001). However, PS was not superior to AS in predicting prognosis (AS vs PS, LR χ²: 16.41 vs 17.5; AIC: 357.44 vs 358.35; C-index, 0.72 vs 0.73, P=0.667). Conclusions Both of AS and PS proposed in the 8th edition American Joint Committee on Cancer (AJCC) staging manual were predictive factors in patients with PMBC. Compared with AS, the PS did not show superiority in prognosis prediction among patients with PMBC.

PURPOSE: The purpose of this study was to investigate the changes of tumor infiltrating lymphocytes (TILs) between core needle biopsy (CNB) and surgery removed sample (SRS) in early stage breast cancer patients and to identify the correlating factors and prognostic significance of TILs changes. MATERIALS AND METHODS: A retrospective study was carried out on 255 patients who received CNB and underwent surgical resection for invasive breast cancer. Stromal TILs levels of CNB and SRS were evaluated respectively. Tumors with ≥50% stromal TILs were defined as lymphocyte-predominant breast cancer (LPBC). Clinicopathological variables were analyzed to determine whether there were factors associated with TILs changes. Log-rank tests and Cox proportional hazards models were used to analyze the influences of TILs and TILs changes on survival. RESULTS: SRS-TILs (median, 10.0%) were significant higher than CNB-TILs (median, 5.0%; p<0.001). Younger age (<60 years, p=0.016) and long surgery time interval (STI, ≥4 days; p=0.003) were independent factors correlating with higher TILs changes. CNB-LPBC patients showed better breast cancer-free interval (BCFI, p=0.021) than CNB-non-LPBC (CNB-nLPBC) patients. Patients were categorized into four groups according to the LPBC change pattern from CNB to SRS: LPBC→LPBC, LPBC→nLPBC, nLPBC→LPBC, and nLPBC→nLPBC, with estimated 5-year BCFI 100%, 100%, 69.7%, and 86.0% (p=0.016). nLPBC→LPBC pattern was an independent prognostic factor of worse BCFI (hazard ratio, 2.19; 95% confidence interval, 1.06 to 4.53; p=0.035) compared with other patterns. CONCLUSION: TILs were significantly higher in SRS than in CNB. Higher TILs changes were associated with younger age and long STI. Changing from nLPBC to LPBC after CNB indicated a worse BCFI, which needs further validation.
Biopsy, Large-Core Needle ; Breast Neoplasms ; Breast ; Humans ; Lymphocytes, Tumor-Infiltrating ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Sexually Transmitted Diseases

Objective: To analyze adjuvant chemotherapy decisions for triple negative breast cancer (TNBC), and explore the influencing factors in the multidisciplinary treatment (MDT) modality. Methods: A retrospective analysis was performed. The cases with invasive TNBC who underwent surgery and MDT discussion for adjuvant treatment in Ruijin Hospital, from April 2013 to June 2015, were recruited. The patients′ clinicopathological characteristics were analyzed and adjuvant treatment suggestions from MDT were obtained. Here the chemotherapy decision alteration was defined as a disagreement in chemotherapy or not, or inconsistence in regimens between the attending doctor and the multidisciplinary team. Results: A total of 194 patients aged ≤70 years old were enrolled in the multidisciplinary discussion, and 187 patients (96.4%) were suggested to receive chemotherapy. When compared the opinions of the attending doctor to suggestions of the multidisciplinary team, we found that the percentage of chemotherapy decision alteration reached 22.7% (39/172), of which 94.9% (37/39) were inconsistence in chemotherapy regimens. There were 119 patients who were recommended to receive epirubicin plus cyclophosphamide (EC) followed by docetaxel (T) or weekly paclitaxel (wP) regimens. Before the announcement of results for the E1199 trial, EC-T accounted for 62.5% (55/88), and EC-wP accounted for 37.5% (33/88) for this group of patients. After that, the proportion of EC-T was decreased to 22.6% (7/31) and proportion of EC-wP increased to 77.4%(24/31) (P<0.001). In addition, a total of 20 patients were suggested to receive platinum based chemotherapy. The proportions were 9.3% in cases with invasive ductal carcinoma, and 33.3% in cases with metaplastic carcinoma, respectively (P=0.016). Conclusions The adjuvant chemotherapy decision for TNBC patients is altered in 22.7% of the patients after MDT discussion. After the announcement of SABCS E1199 results, more patients are suggested to receive EC followed by weekly paclitaxel. There is a lack of detailed evidence for platinum based adjuvant chemotherapy for TNBC, and more patients with metaplastic carcinoma receive platinum based adjuvant chemotherapy.

Objective: To evaluate the choice of surgical treatment of ductal carcinoma in situ (DCIS) and its impact on long-term outcomes. Methods: A retrospective analysis of the clinicopathological features and treatment protocol of DCIS patients who underwent surgical treatment in Comprehensive Breast Health Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine from January 2009 to August 2016 was done. The factors which could affect surgical treatment were analyzed by χ² test and Logistic regression. Survival analysis were performed between different surgical approaches. Kaplan-Meier survival curves and Log-rank tests demonstrated the distribution of disease free survival and overall survival. Results: A total of 526 patients were enrolled in this study, 405 cases (77.0%) underwent mastectomy, 121 cases (23.0%) underwent breast-conserving surgery, of which 88 cases received radiotherapy after breast-conserving surgery. It was shown by univariate and multivariate analysis that age>50 years (OR=0.631, 95% CI: 0.413 to 0.965, P=0.034), first symptom of nipple discharge (OR=0.316, 95% CI: 0.120 to 0.834, P=0.020), excision biopsy (OR=1.831, 95% CI: 1.182 to 2.835, P=0.007) and tumor size >3 cm (OR=0.422, 95% CI: 0.206 to 0.864, P=0.018) were significantly correlated with choice of surgical treatment for breast lesions. Axillary lymph node dissection was performed for 118 cases (22.4%), with sentinel lymph node biopsy for 327 cases (62.2%), and none for 81 cases (15.4%). There was significant statistical difference in the choice of axillary lymph node management in patients of different age (χ²=8.124, P=0.017), biopsy type (χ²=35.567, P=0.000), breast operation type (χ²=149.118, P=0.000) and tumor size (χ²=13.394, P=0.010). The 5-year disease free survival rates was 95.7%, 89.6% and 100%, respectively, for mastectomy group, breast-conserving surgery group and breast-conserving surgery plus radiotherapy group. And the 5-year overall survival rates for three groups were 99.0%, 100% and 100%. The differences were not statistically significant (P=0.427, 0.777). Conclusions For DCIS patients, age, first symptom and tumor size are independent predictors of breast surgery. The choice of axillary lymph node surgery is influenced by age, biopsy, operation type, and tumor size. Different surgical treatment options has no significant effect on disease-free survival and overall survival in DCIS patients.

Objective: To investigate the effect of 21-gene recurrence score on adjuvant chemotherapy decisions for patients with estrogen receptor (ER)-positive, epidermal growth factor receptor 2 (HER-2)-negative and lymph node (LN)-negative early stage-breast cancer. Methods: One hundred and forty-eight patients with ER+ , HER-2- and LN- early stage breast cancer were recruited in the Ruijin hospital, Shanghai Jiao Tong University School of Medicine. The 21-gene recurrence score (RS)assay was performed and systemic therapeutic decisions were made before and after knowing the RS results under multidisciplinary discussion. The effects of RS assay and the other influential factors on adjuvant chemotherapy decision were further analyzed. Results: After knowing the RS results, treatment decisions were changed in 26 out of 148 patients(17.6%). Among them, 9 out of 26 patients were not recommended for chemotherapy; 16 of 26 had treatment recommendation changed to chemotherapy, and chemotherapy regimen was changed in the last one patient. Multivariate analysis showed that RS, age and histological grade were independent factors of decision-making for adjuvant chemotherapy. Conclusion Our results suggest that 21-gene recurrence score significantly influences decision making for adjuvant chemotherapy in patients with ER+ , HER-2- and LN- early stage breast cancer.

Objective: To investigate the distribution patterns of 21-gene assay and its influencing factors in Chinese patients with early breast cancer. Methods: Nine hundred and twenty-seven early breast cancer patients were retrospectively recruited from January 2009 to December 2015 at Ruijin Hospital, Shanghai Jiaotong University School of Medicine. The 21-gene reverse transcriptase-polymerase chain reaction(RT-PCR) assay were conducted in paraffin-embedded tumor tissues to calculate the Recurrence Score(RS). Immunohistochemistry(IHC) assay was used to measure the expression levels of estrogen receptor(ER), progesterone receptor(PR) and Ki-67. Concordances of RT-PCR and IHC results were assessed. Correlations of RS and classical clinicopathological factors were evaluated, and logistic regression were applied to determine independent predictive factors for RS. Results: The median RS of 927 patients was 23(range: 0~90), and the proportions of patients categorized as having a low, intermediate, or high risk were 26.5%, 47.7% and 25.8%, respectively. The distribution of RS varied significantly according to different tumor grade, T stage, PR status, Ki-67 index and molecular subtypes(P<0.05 for all). Grade, PR status and Ki-67 index were independent predictive factors for RS. ER, PR status and Ki-67 index showed significantly correlation between RT-PCR and IHC assays, and the concordance rates for ER and PR status were 98.7% and 87.8%, respectively. Conclusions RS significantly correlated with tumor grade, T stage, PR status, Ki-67 index and subtypes. Grade, PR status and Ki-67 index can independently predict RS. Remarkable concordances of ER, PR status and Ki-67 index are found between RT-PCR and IHC assays.