Tertiary lymphoid structures in gynecological cancers: prognostic role, methods for evaluating, antitumor immunity, and induction for therapy.

Tertiary lymphoid structures (TLSs), referred to as tertiary lymphoid organs and lymphoid tissue neogenesis, are aggregates of immune cells that occur in nonlymphoid tissues. In recent years, it has been found that TLSs within the tumor microenvironment have been associated with local adaptive immune immunity against cancer and favorable prognosis in several human solid tumors, including gynecological cancers. The issue of the prognosis of gynecological cancers, including endometrial, cervical, and ovarian cancer, is an enormous challenge that many clinical doctors and researchers are now facing. Concerning the predictive prognostic role of TLSs, effective evaluation, and quantification of TLSs in human tissues may be used to assist gynecologists in assessing the clinical outcome of gynecological cancer patients. This review summarizes the current knowledge of TLSs in gynecological cancers, mainly focusing on the potential mechanism of TLS neogenesis, methods for evaluating TLSs, their prognostic value, and their role in antitumor immune immunity. This review also discusses the new therapeutic methods currently being explored in gynecological cancers to induce the formation of TLSs.

A preoperative nomogram predicting risk of lymph node metastasis for early-stage cervical cancer.

OBJECTIVE: This study aimed to develop a preoperative nomogram based on clinical and pathological characteristics to provide a more individualized and accurate estimation of lymph node metastasis (LNM) in patients with early-stage cervical cancer. METHODS: A total of 7,349 early-stage cervical cancer patients with pathologically confirmed between 1988 and 2015 were obtained from the Surveillance, Epidemiology, and End Results (SEER) database. All the patients were divided into training (n = 5,500) and validation (n = 1,849) cohorts randomly. A cohort of 455 patients from multicenter was used for the external validation. We established a multivariate logistic regression model based on preoperative clinicopathological data, from which a nomogram was developed and validated. A predicted probability of LNM < 5% was defined as low risk. RESULTS: From multivariate logistic regression analysis, age at diagnosis, histologic subtype, tumor grade, tumor size and FIGO stage were identified as preoperative independent risk factors of LNM. The nomogram incorporating these factors demonstrated good discrimination and calibration (concordance index = 0.723; 95% confidence interval (CI), 0.707-0.738). In the validation cohort, the discrimination accuracy was 0.745 (95% CI, 0.720-0.770) and 0.747 (95% CI, 0.690-0.804), respectively. The nomogram was well calibrated with a high concordance probability. We also established an R-enabled Internet browser for LNM risk assessment, which tool may be convenient for physicians. CONCLUSIONS: We developed an effective preoperative nomogram based on clinical and pathological characteristics to predict LNM for early-stage cervical cancer. This model could improve clinical trial design and help physicians to decide whether to perform lymphadenectomy or not.

Tertiary Lymphoid Structures Are Associated with a Favorable Prognosis in High-Grade Serous Ovarian Cancer Patients.

There was accumulating evidence indicating that tertiary lymphoid structures (TLSs) were strongly associated with improved survival and clinical outcome in several solid tumors. In this study, we intended to assess the presence of TLSs and their potential clinical significance in high-grade serous ovarian cancer (HGSOC). TCGA (The Cancer Genome Atlas) cohort included RNA-seq data of 376 HGSOC patients, of which 74 patients included available hematoxylin-eosin (H&E) sections; GEO (Gene Expression Omnibus) cohort, GSE140082, included microarray data of 212 HGSOC patients. TLSs were counted by pathological sections, and the relative abundance of TLSs was assessed by the unsupervised consensus clustering of 12-chemokine transcriptome signatures. The potential associations between TLSs and clinical prognosis, tumor microenvironment (TME), and immunotherapy response of HGSOC were further performed based on transcriptome data. In the H&E sections of HGSOC, TLSs were predominantly located in the stroma and invasive margin of the tumor. Pathological counting results suggested that the expression of 12 chemokines was significantly higher in samples abundant with TLSs than that in the lack of TLSs. Consensus clustering of both TCGA and GEO cohorts divided HGSOC patients into two clusters with different TLSs abundance: low- and high-TLSs. Based on transcriptome analysis, the high-TLS cluster was characterized by better clinical prognosis, a higher degree of immune infiltration, more biological pathways, higher tumor mutational burden score, and higher expression of immune checkpoints. In conclusion, TLSs strongly correlated with the immune-responsive microenvironment and remained a favorable prognostic factor independent of other clinical characteristics in HGSOC. The presence of TLSs was also associated with a potentially favorable response to immune checkpoint blockade (ICB) therapy in HGSOC.

ZEB1 promotes invasion and metastasis of endometrial cancer by interacting with HDGF and inducing its transcription.

Zinc finger E-box binding homeobox 1 (ZEB1), as a typical transcription inhibitory factor of E-cadherin, plays a major role in stimulating the invasion and metastasis of tumors via modulating the epithelial-mesenchymal transition (EMT) signal. However, its function and modulatory mechanisms in endometrial carcinoma (EC) remain unclear. In this study, silencing ZEB1 significantly reduced EC cell migration, invasion, and metastasis, as well as enhanced the sensitivity of EC cells to cisplatin (cDDP) in vitro and in vivo. Mechanism analysis indicated that ZEB1 interacts with hepatoma-derived growth factor (HDGF) and co-localizes in the nucleus. In addition, ZEB1 as a transcription factor binds to the promoter of HDGF to stimulate HDGF transcription. Furthermore, suppression of HDGF in ZEB1-overexpressed EC cells not only reduced the expression of ß-catenin, TCF4, and ZEB1, but also repressed ß-catenin translocation from the cytoplasm into the nucleus and further downregulated the combination with TCF4. Interestingly, the ß-catenin/TCF4 signaling feedback stimulates ZEB1 transcription and therefore constitutes a positive feedback loop. In clinical samples, ZEB1 positively correlates with HDGF expression, and co-expression of ZEB1 and HDGF promotes the pathogenesis of EC. In summary, our study demonstrated that the positive feedback loop of ZEB1/HDGF/ß-catenin/TCF4 plays an unfavorable role in the metastasis of endometrial carcinoma.

Sp1 contributes to radioresistance of cervical cancer through targeting G2/M cell cycle checkpoint CDK1.

BACKGROUND/AIMS: Radioresistance remains a significant obstacle in the therapy of cervical cancer, and the mechanism of it is still unclear. We aimed to investigate the role of specificity protein 1 (Sp1) in radioresistance of cervical cancer. METHODS: Sp1 was examined immunohistochemically on tissues from 36 human cervical cancer patients. We used RT-qPCR and Western blot to examine the expression of Sp1 in irradiated cervical cancer cell lines SiHa and HeLa. The role of Sp1 in radioresistance of cervical cancer cells was assessed by colony-formation assay and cell cycle analysis. Dual-luciferase reporter assay was performed to detect the downstream of Sp1. RESULTS: High Sp1 expression was positively correlated with advanced International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis, and lymphovascular space invasion (LVSI) of cervical cancer. The expression of Sp1 was dose-dependently increased in irradiated cervical cancer cell lines at both mRNA and protein levels. Colony-formation assay showed that alteration of Sp1 expression affected the survival of cervical cancer cells with radiotherapy (RT) treatment. Knockdown of Sp1 significantly strengthened the cellular response to radiation by inducing G2/M arrest in cervical cancer cells. Overexpression of Sp1 significantly decreased G2/M arrest in cervical cancer cells, which was related to upregulation of CDK1 expression. Dual-luciferase reporter assay showed the direct effect of Sp1 on the transcriptional activation of CDK1. CONCLUSION: Sp1 may contribute to radioresistance through inhibiting G2/M phase arrest by targeting CDK1, and be considered as a potential therapeutic target to promote the effect of RT for patients with cervical cancer.

[Expression of phosphoglycerate kinase 1 in endometrial carcinoma and its association with patients' outcome].

OBJECTIVE: To investigate the expression of phosphoglycerate kinase 1 (PGK1) and its prognostic value in endometrial carcinoma (EC). METHODS: The expression of PGK1 was detected immunohistochemically in 30 normal endometrium and 130 EC specimens. The relationship between PGK1 protein expression and the clinicopathological features of the patients was evaluated. RESULTS: Immunohistochemical analysis revealed low PGK1 expression in 55.4% (72/130) and high PGK1 expression in 44.6% (58/130) of the EC specimens, as compared with the rates of 90% (27/30) and 10% (3/30) in normal endometrium, respectively (P<0.001). PGK1 expression was significantly correlated with FIGO stage (P<0.001), histological grade (P=0.002) and lymph node metastasis (P<0.001). Kaplan-Meier survival analysis indicated that patients with a high PGK1 expression had a shorter overall survival rate than those with a low PGK1 expression (P<0.001). Multivariate analysis showed that a high PGK1 expression was not the independent predictor of the prognosis of EC (P=0.077). CONCLUSION: A high expression of PGK1 is associated with aggressive and metastatic behaviors of EC, and detection of PGK1 provides assistance in evaluating the prognosis of patients with EC.

[Role of specificity protein 1 in modulating radiosensitivity of cervical cancer cell lines].

OBJECTIVE: To investigate the role of specificity protein 1 (Sp1) in regulating radiosensitivity of cervical cancer cell lines. METHODS: We analyzed Sp1 expression in 6 different cervical cancer cell lines (SiHa, HeLa, Caski, Me180, Ms751, and C33a) using Western blotting and real-time PCR. Clonogenic survival assay and curve fitting were used to assess the changes in radiosensitivity of Me180 cells transfected with lentivirus-mediated shRNA vector targeting sp1 and HeLa cells transfected with sp1 over-expression vector. RESULTS: In the 6 cell lines tested, the cellular expression levels of Sp1 decreased gradually in the order of Me180, Caski, C33a, SiHa, Ms751, and HeLa. SP1 knockdown with lentivirus-mediated shRNA significantly lowered the survival rate of Me180 cells following radiation exposure (P<0.05), and obviously lowered the values of SF2, D0 and Dq but significantly increased α/ß of the cells. Compared with the cells transfected with the mock vector, HeLa cells with sp1 over-expression showed a significantly increased survival following radiation exposure (P<0.05) with obviously increased values of SF2, D0 and Dq but significantly lowered α/ß. CONCLUSION: Silencing Sp1 can increase the radiosensitivity while Sp1 overexpression enhances the radioresistance of cervical cancer cell lines, suggesting an important role of Sp1 in radiotherapy for cervical cancer.

[Expressions of DAPK3 and c-Myc in endometrial cancer and their relationship with the patients' prognosis].

OBJECTIVE: To investigate the expressions of DAPK3 and c-Myc and their prognostic value in endometrial carcinoma (EC). METHODS: The expressions of DAPK3 and c-Myc were detected immunohistochemically in 132 surgical specimens. The relationship between DAPK3 and c-Myc protein expressions and the clinicopathological features of the patients was evaluated. RESULTS: Immunohistochemical analysis revealed low DAPK3 expression in 60.61% (80/132) and high c-Myc expression in 53.79% (71/132) of the specimens of EC. Both DAPK3 expression and c-Myc expression were significantly correlated with FIGO stage (P=0.034 and 0.015, respectively) and lymph node metastasis (P=0.022 and 0.031, respectively). DAPK3 expression was closely correlated with the histological grade (P=0.027) and depth of myometrial invasion (P=0.011). Kaplan-Meier survival analysis indicated that patients with low DAPK3 expressions had a shorter overall survival rate than those with high DAPK3 expressions (P=0.023), while patients with high c-Myc expressions had poorer prognoses than those with low c-Myc expressions (P=0.002). Spearman correlation analysis showed that DAPK3 and c-Myc expressions were negatively correlated (P<0.001, r=?0.310). Multivariate analysis identified a high c-Myc expression as the independent predictor of the prognosis of EC (P=0.007). CONCLUSIONS: A low expression of DAPK3 and a high expression of c-Myc are associated with aggressive and metastatic behaviors of EC, and their detection may help to predict the prognosis of the EC patients.

[Coexpression of MAP2K4 and vimentin proteins in human endometrial carcinoma and its clinicopathological significance].

OBJECTIVE: To analyze the expression of MAP2K4 and vimentin in human endometrial carcinoma (EC) and their association with the clinicopathological features and prognosis of the patients. METHODS: MAP2K4 and vimentin expressions were detected immunohistochemically in paraffin-embedded tissue sections from 128 patients with EC, and the correlation of MAP2K4 and vimentin expressions with the clinicopathological factors of the patients was analyzed. RESULTS: MAP2K4 and vimentin proteins were positively expressed in 49 (38.3%) and 83 (64.8%) of the patients, respectively. A positive expression of MAP2K4 was negatively correlated with FIGO stage of the tumor (P=0.010) and lymph node status (P=0.016); a positive expression of vimentin was positively correlated with FIGO stage of the tumor (P=0.025), histological grades (P=0.017), depth of myometrial invasion (P=0.044) and lymph node status (P=0.032). MAP2K4 was inversely associated with vimentin expression in EC(r=-0.598, P<0.001). Patients positive for MAP2K4 tended to have a higher overall survival rate (P=0.002), and those positive for vimentin tended to have a lower overall survival rate (P=0.007); patients positive for MAP2K4 but negative for vimentin had the longest survival time, while those negative for MAP2K4 and positive for vimentin had lowest survival rate (P=0.004). CONCLUSION: Detection of MAP2K4 and vimentin might help in early diagnosis and prognostic evaluation of patients with EC.

[Event-related potentials of emotional excitement in middle-aged and aged adults].

OBJECTIVE: To explore how adjustment of the negative and neutral emotion excitement contribute to the well-being of middle-aged and aged adults in their health, intelligence, working and living. METHODS: Twenty-five middle-aged and aged participants performed recognition tasks of schematic facial representations of positive, neutral and negative emotions and also Chinese characters (equivalent to happy, thinking and fear). RESULTS: The main effects of the facial recognition task occurred in the left temporal-parietal area (40 to 80 ms), bilateral parietal-occipital and temporal-parietal area (160-180 ms and 310-340 ms). The effects of stimulation with the 3 Chinese characters were found in the anterior region (375-475 ms), posterior region (195-255 ms), and whole scalp region (135-175 ms and 275-355 ms). There were no significant interactive effects between schematic facial recognition and stimulation with Chinese characters. CONCLUSION: Positive emotional excitement may promote the well-being of the adults in their health, intelligence, working performance and living.

[Preliminary study of gene expression profiling in human type I and II endometrial carcinoma].

OBJECTIVE: To study gene expression profiling in human type I and II endometrial carcinoma. METHODS: Six Affymetrix human genome genechips were utilized to investigate the differences in gene expression profiles between type I and II endometrial carcinoma with bioinformatic analysis. RESULTS: Many genes were highly expressed in estrogen-dependent endometrial carcinoma, and some of them were involved in the metabolism and conversion of estrogen, while some others in estrogen regulation. CYP2C9, for instance, was involved in the conversion of estrogen sulfate to 16-hydroxy sulfate metabolite, DDC in estrogen-dependent pathogenesis of endometrial carcinoma possibly by DDC interaction with AR to enhance steroid receptor transcription. CONCLUSION: High expression of these genes in estrogen-dependent endometrial carcinoma may provide insights into their roles in the pathogenesis and prognosis of this malignancy.

[Detection of human papillomavirus DNA in cervical cancer tissue by PCR and direct sequencing].

OBJECTIVE: To explore an effective method for detecting human papillomavirus (HPV) DNA in cervical cancer tissue. METHODS: HPV L1 gene fragment in cervical cancer tissue was amplified by HPV-specific PCR with consensus primers, and typing of the HPV strains was performed on the basis of sequence analysis of the PCR product. RESULTS: The positivity rates of HPV DNA was 78% in the 50 cases of cervical cancer, and mixed infection with HPV16 and HPV18 strains was the most common, which accounted for 48% on the total infections. Infection with HPV58 was detected in one case. The sequencing results showed no difference in L1 sequence between the detected samples and the standard German HPV58 strain. CONCLUSION: PCR and direct sequencing approach is effective for detecting and typing of HPV DNA in cervical cancer tissue, through which rare HPV strain or mutants of known HPV strains may not escape detection.