Microvasculature and microstructure alteration in dry-type high myopia.

BACKGROUND: To investigate the alterations in the retinal vasculature and microstructure in dry-type high myopia. METHODS: One hundred and eighty-nine dry-type high myopia eyes were classified into three groups. Group 1 consisted of 86 eyes with no myopic retinal degenerative lesion (C0). Group 2 consisted of 71 eyes with tessellated fundus (C1). Group 3 consisted of 32 eyes with diffuse chorioretinal atrophy (C2). Retinal vascular density and retinal thickness were measured with optical coherence tomography angiography. The scanning area was a 3 × 3 mm2 ring with the fovea of the macular. All data were analyzed with the SPSS 23.0 by one-way ANOVA test among comparison groups. Pearson's correlation analysis was used to determine the relations among measurements. Univariate linear regression showed a correlation between the vascular densities and retinal thicknesses. RESULTS: The microvessel density significantly decreased and significant thinning of the superior and temporal macular thickness in the C2 group. The vascular densities of macular decreased significantly with the increase of axial length (AL) and refractive diopter in the C2 group. The retinal thicknesses of the macular fovea increased significantly with the increase of vascular densities in the C0 group and C1 group. CONCLUSIONS: The impairment of retinal microstructure is more likely related to reduced oxygen and nutrients due to microvessel density decreases.

Novel role of PAF1 in attenuating radiosensitivity in cervical cancer by inhibiting IER5 transcription.

BACKGROUND: Radiosensitivity is limited in cervical cancer (CC) patients due to acquired radiation resistance. In our previous studies, we found that immediate-early response 5 (IER5) is upregulated in CC cells upon radiation exposure and decreases cell survival by promoting apoptosis. The details on the transcriptional regulation of radiation-induced IER5 expression are unknown. Studies in recent years have suggested that Pol II-associated factor 1 (PAF1) is a pivotal transcription factor for certain genes "induced" during tumor progression. In this study, we investigated the role of PAF1 in regulating IER5 expression during CC radiotherapy. METHODS: PAF1 expression in CC cells was measured by western blotting, immunohistochemistry, and qRT-PCR, and the localization of PAF1 and IER5 was determined by immunofluorescence. The effect of PAF1 and IER5 knockdown by siRNA in Siha and Hela cells was studied by western blotting, qRT-PCR, CCK-8 assay, and flow cytometry. The physical interaction of PAF1 with the IER5 promoter and enhancers was confirmed using chromatin immunoprecipitation and qPCR with or without enhancers knockout by CRISPR/Cas9. RESULTS: We confirmed that PAF1 was highly expressed in CC cells and that relatively low expression of IER5 was observed in cells with highly expressed PAF1 in the nucleus. PAF1 knockdown in Siha and Hela cells was associated with increased expression of IER5, reduced cell viability and higher apoptosis rate in response to radiation exposure, while simultaneous PAF1 and IER5 knockdown had little effect on the proportion of apoptotic cells. We also found that PAF1 hindered the transcription of IER5 by promoting Pol II pausing at the promoter-proximal region, which was primarily due to the binding of PAF1 at the enhancers. CONCLUSIONS: PAF1 reduces CC radiosensitivity by inhibiting IER5 transcription, at least in part by regulating its enhancers. PAF1 might be a potential therapeutic target for overcoming radiation resistance in CC patients.

Clinical significance and functions of microRNA-93/CDKN1A axis in human cervical cancer.

AIM: Accumulating studies have revealed that microRNA (miR)-93 may exert an oncogenic role in various cancers via inhibiting CDKN1A. However, the involvement of miR-93/CDKN1A axis in cervical cancer remains unclear. We aimed to investigate expression pattern, clinical significance and potential functions of miR-93/CDKN1A axis in cervical cancer. METHODS: Expression levels of miR-93 and CDKN1A mRNA in 100 pairs of cervical cancer and matched non-cancerous tissue samples were detected by quantitative-PCR. Statistical analyses were performed to evaluate associations of miR-93 and/or CDKN1A expression with various clinicopathologic features and patients' prognosis. The functions of miR-93/CDKN1A axis on cell proliferation and invasion were also examined. RESULTS: Compared to non-cancerous tissues, the expression levels of miR-93 and CDKN1A were dramatically increased and decreased in cervical cancer tissues, respectively (both P < 0.01). High miR-93 and/or low CDKN1A expression were significantly associated with advanced International Federation of Gynecology and Obstetrics stage, the presence of lymph node metastasis and recurrence (all P < 0.05). Importantly, patients with high miR-93 and/or low CDKN1A expression had shorter overall survival than those with low miR-93 and/or high CDKN1A expression. The multivariate analysis identified miR-93 and/or CDKN1A expression as independent prognostic factors of cervical cancer. Functionally, miR-93 promoted cell proliferation and invasion of cervical cancer cells via inhibiting CDKN1A. CONCLUSION: miR-93 upregulation and CDKN1A downregulation may be both associated with the development, progression and patients' prognosis of cervical cancer. miR-93/CDKN1A axis may also play an important role in the malignancy of cervical cancer cells, suggesting its potential as a therapeutic target for this cancer.

Incidence of radiation enteritis in cervical cancer patients treated with definitive radiotherapy versus adjuvant radiotherapy.

OBJECTIVE: The study aimed to evaluate the incidence of early and late radiation enteritis in cervical cancer patients receiving definitive or adjuvant radiotherapy (RT). MATERIALS AND METHODS: Three hundred and twenty-four cervical cancer patients receiving definite or adjuvant RT in our hospital from January 2010 to December 2012 were divided into definitive (132 patients) or adjuvant RT (192 patients) and performed detailed analysis. RESULTS: Early radiation enteritis was found in 54.3%, and late radiation enteritis was found in 17.9% of cervical patients. Early (P < 0.007) and late (P < 0.003) radiation enteritis appeared more frequently in patients treated with definitive RT than that with adjuvant RT. In the definitive RT group, incidence of both early and late radiation enteritis in the RT group was higher than in the radiochemotherapy (RCHT) group, and the difference was statistically significant (P < 0.004). Severe cases of late radiation enteritis (Grade 3 and 4) can be seen higher in the definitive radiation group (both the RT and RCHT group) than in the adjuvant radiation group, and the difference was statistically significant (P < 0.005). CONCLUSION: The incidence of both early and late radiation enteritis in the definitive RT group is higher than in the adjuvant RT group. The occurrence of side effects was associated with the prolongation of total irradiation time due to necessary interruptions of RT. Methods to decrease the interruptions in the RT and the irradiated volume of the small bowel will further lessen enteric morbidity.

Clinical efficacy of preoperative vaginal intracavitary irradiation for Stage Ib2 and IIa cervical cancer.

OBJECTIVE: The purpose of this study was to evaluate the clinical efficacy of preoperative vaginal intracavitary irradiation for Stage Ib2 and IIa cervical cancer. MATERIALS AND METHODS: From June 2008 to June 2014, data from 78 Stage Ib2 and IIa cervical cancer patients (age ≤75 years) with a diameter of local lesions >4 cm were collected in our hospital. Before treatment, all cases were confirmed by biopsy. The patients' general state was good, Karnofsky Performance Score ≥90, heart and lung functioning was normal, and patients were able to tolerate the surgery. The 78 patients were randomly divided into two groups: neoadjuvant radiotherapy group (NRG) (n = 38) and radical surgery group (n = 40). Patients in NRG received 2000~3000 cGy192 Ir irradiation of preoperatively intracavitary brachytherapy (radioactive source at 1 cm distance). After a rest of 10-14 days, radical hysterectomy with pelvic lymphadenectomy was performed. Surgery alone group (SAG) (n = 40) underwent radical surgery directly. The treatment outcomes between these two groups were compared, and the effect of preoperative intracavitary brachytherapy and presence of postoperative complications were evaluated. RESULTS: The total clinical efficacy for intracavitary brachytherapy was 94.7% (36/38) with complete response 13 (34.2%), partial response 23 (60.5%), and stable disease 2 (5.3%). Moreover, no patients developed progression disease; for SAG patients, 32 cases successfully finished the extensive hysterectomy and pelvic lymphadenectomy. Operation time <240 min was found in 19 patients. Moreover, the positive complication for lymphatic cyst and urinary retention was 20.0% and 15.0%, respectively. For NAG group, 36 patients successfully finished the extensive hysterectomy and pelvic lymphadenectomy. Operation time <240 min was found in 22 patients. Moreover, the positive complication for lymphatic cyst and urinary retention was 15.8% and 13.2%, respectively. The median follow-up time for NRG and SAG was 28 and 30 months, respectively. Three cases lost to follow-up in the SAG with the follow-up rate of 92.5% (37/40). In the NRG group, 3 cases lost to follow-up with the follow-up rate of 92.1% (35/38). The locoregional control rate for 1, 3, and 5 years was 80.0%, 61.3%, and 52.6%, respectively, for SAG group and 89.5%, 82.9%, and 76.9%, respectively, for NRG group with significant statistical difference for 3 and 5 years. CONCLUSION: Preoperative intracavitary brachytherapy is an effective procedure for the treatment for Stage Ib2 and IIa cervical cancer and can significantly improve the locoregional control rate.

IER5 as a promising predictive marker promotes irradiation-induced apoptosis in cervical cancer tissues from patients undergoing chemoradiotherapy.

PURPOSE: To define the role of immediate-early 5 (IER5) gene as a promising biomarker in predicting the radiosensitivity and prognosis of cervical cancer patients receiving cisplatin-based concurrent chemoradiotherapy (DDP-CCRT). RESULTS: Our investigations found that IER5 level was markedly elevated in cervical cancer patients after being treated with irradiation, which indicated IER5 was closely dose induced. By contrast, the correlation between IER5 and radiosensitivity cannot be confirmed by the present study. The up-regulation of IER5 expression effectively increased cell apoptosis after administration of irradiation (P < 0.05). Using an ANOVA model for repeated-measures, we found significant association between the IER5 level and tumor size (P < 0.05). MATERIALS AND METHODS: Forty-three cervical cancer patients stage IIb-IIIb received DDP-CCRT were registered. Biopsy tissues were obtained after administration of irradiation dose of 0 Gy, 2~6 Gy, 10 Gy, 20 Gy, 30 Gy, respectively. The IER5 protein and mRNA levels were measured by immunohistochemistry, western blot and quantitative polymerase chain reaction, respectively; besides, the apoptosis rate was assessed by transferase-mediated dUTP nick end labeling. CONCLUSIONS: Mechanistically, we confirmed that IER5 induced by radiation dose enhanced apoptosis of cervical cancer, was inversely associated with tumor size. In conclusion, our studies indicate target IER5 is improved to be a potential radiosensitizer for developing effective therapeutic strategies against cervical cancer to radiotherapy and a predictive biomarker for radiosensitivity.

Fyn-related kinase expression predicts favorable prognosis in patients with cervical cancer and suppresses malignant progression by regulating migration and invasion.

AIM: To investigate expression pattern, clinical significance and potential roles of fyn related kinase (FRK) in cervical cancer. METHODS: Expression of FRK protein and mRNA in 100 pairs of cervical cancer and matched non-cancerous tissue samples were detected by Western blot, immunohistochemistry and quantitative PCR, respectively. Statistical analyses were performed to evaluate associations of FRK protein expression with various clinicopathologic features and patients' prognosis. The effects of FRK on cell migration and invasion were examined using in vitro migration and invasion assays, respectively. RESULTS: Weak/negative immunostaining of FRK protein was observed in 86 (86.00%) of 100 cervical cancer tissues. Low FRK expression was significantly associated with several aggressive clinicopathologic features of cervical cancer, such as higher International Federation of Gynecology and Obstetric stage (P=0.01), the presence of lymph node metastasis (P=0.01) and recurrence (P=0.02). In addition, the survival analysis showed that cervical cancer patients with low FRK expression often had shorter overall survival than those with high FRK expression. The multivariate analysis also identified FRK expression as an independent prognostic factor of cervical cancer. Functionally, the enforced expression of FRK could efficiently inhibit cell migration and invasion of cervical cancer cells, but the knockdown of FRK dramatically enhanced cell migration and invasion. CONCLUSION: Our findings suggest that loss of FRK protein may be implicated into the tumorigenesis and cell motility of human cervical cancer. More importantly, FRK expression may function as a promising prognostic marker of this malignancy, highlighting its potentials as a candidate target for gene therapy.

Effect of preoperative radiotherapy on stage IB2 and IIA2 cervical cancer: A retrospective cohort study.

INTRODUCTION: The aim of the retrospective study was to investigate the therapeutic efficiency of the preoperative intracavitary radiotherapy combined with radical surgery on postoperative complications and long-term survival in patients with stage IB2 and IIA2 cervical cancer (CC). METHODS: From January 1995 to December 2012, a total of 171 patients with stage IB2 or IIA2 CC were recruited into the study. They were divided into two groups according to the treatment modality provided: preoperative radiotherapy followed by radical surgery (n = 80), and radical surgery alone (n = 91). The clinical curative effect, postoperative complications and the postoperative prognosis of patients were evaluated and compared in two groups. The tumor response and survival of patients in two groups were observed in follow-up study. RESULTS: There were no significant differences in the incidence of postoperative complications, intraoperative blood loss and surgery duration (P > 0.05) between the two groups. Preoperative radiotherapy did not improve the postoperative prognosis yet. Though patients undergoing preoperative radiotherapy showed the similar 1- (92.50% vs. 84.62%), 3- (85.00% vs. 81.32%) and 5-year (80.00% vs. 74.72%) survival rates, the 3- and 5-year locoregional control rates of them were much higher than those undergoing surgery alone (P < 0.05). CONCLUSION: Preoperative radiotherapy combined with radical surgery could improve locoregional control rate and would not increase the risk of postoperative complications. It may be a feasible treatment mode for early stage CC carcinoma.

Identification of DNA methylation of SOX9 in cervical cancer using methylated-CpG island recovery assay.

The aim of the present study was to identify novel methylation markers for cervical cancer screening and to test the clinical application of the most promising biomarker in cervical scrapings. Methylated-CpG island recovery assay-based microarray analysis was carried out on a discovery set consisting of cervical cancer tissue and normal cervical tissue to identify significantly hypermethylated genes. Five hundred and four CpG islands, corresponding to 378 genes, were differentially methylated between cervical cancer tissue and normal cervical tissue. Among them, 30 genes were significantly hypermethylated. Of the 30 genes, SOX9, PKLR and DLX4 were selected for further validation by direct bisulfite sequencing. The SOX9 gene revealed complete methylation in the cervical cancer tissue and complete non-methylation in the normal control tissue. A TaqMan-based real-time PCR assay was performed to detect the methylation levels of the SOX9 gene in 156 cervical scrapings, including 48 normal cervical scrapings, 30 scrapings with cervical intraepithelial neoplasia 1 (CIN1), 30 scrapings with CIN2-3 and 48 scrapings with squamous cell carcinoma (SCC). The methylation levels (methylation score) of the SOX9 gene increased significantly with the severity of cervical squamous lesions. The area under the receiver operating characteristic (ROC) curve (AUC) revealed that the methylation score of the SOX9 gene could be used to segregate SCC/CIN2-3 from CIN1/normal (AUC, 0.961; p=0.000). At the optimal cut-off value, a sensitivity of 92.3% and a specificity of 89.7% were obtained. In conclusion, SOX9 methylation is frequently involved in cervical carcinogenesis, and may provide a valuable molecular biomarker for early detection of cervical cancer.