Identification of miR-23a as a novel microRNA normalizer for relative quantification in human uterine cervical tissues

Shen, Yuanming,Li, Yang,Ye, Feng,Wang, Fenfen,Wan, Xiaoyun,Lu, Weiguo,Xie, Xing Korean Society for Biochemistry and Molecular Bion 2011 Experimental and molecular medicine Vol.43 No.6

Quantitative real-time RT-PCR (RT-qPCR) is being widely used in microRNA expression research. However, few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in microRNA RT-qPCR studies. The aim of this study was to identify the most stable reference gene(s) for quantification of microRNA expression analysis in uterine cervical tissues. A microarray was performed on 6 pairs of uterine cervical tissues to identify the candidate reference genes. The stability of candidate reference genes was assessed by RT-qPCR in 23 pairs of uterine cervical tissues. The identified most stable reference genes were further validated in other cohort of 108 clinical uterine cervical samples: (HR-HPV- normal, n = 21; HR-HPV+ normal, n = 19; cervical intraepithelial neoplasia [CIN], n = 47; cancer, n = 21), and the effects of normalizers on the relative quantity of target miR-424 were assessed. In the array experiment, miR-26a, miR-23a, miR-200c, let-7a, and miR-1979 were identified as candidate reference genes for subsequent validation. MiR-23a was identified as the most reliable reference gene followed by miR-191. The use of miR-23a and miR-191 to normalize expression data enabled detection of a significant deregulation of miR-424 between normal, CIN and cancer tissue. Our results suggested that miR-23a and miR-191 are the optimal reference microRNAs that can be used for normalization in profiling studies of cervical tissues; miR-23a is a novel microRNA normalizer.

Identification of miR-23a as a novel microRNA normalizer for relative quantification in human uterine cervical tissues

Yuanming Shen,Xing Xie,Yang Li,Feng Ye,Fenfen Wang,Xiaoyun Wan,Weiguo Lu 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.6

Quantitative real-time RT-PCR (RT-qPCR) is being widely used in microRNA expression research. However,few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in microRNA RT-qPCR studies. The aim of this study was to identify the most stable reference gene(s) for quantification of microRNA expression analysis in uterine cervical tissues. A microarray was performed on 6 pairs of uterine cervical tissues to identify the candidate reference genes. The stability of candidate reference genes was assessed by RT-qPCR in 23 pairs of uterine cervical tissues. The identified most stable reference genes were further validated in other cohort of 108 clinical uterine cervical samples:(HR-HPV- normal, n = 21; HR-HPV+ normal, n = 19; cervical intraepithelial neoplasia [CIN], n = 47; cancer, n = 21), and the effects of normalizers on the relative quantity of target miR-424 were assessed. In the array experiment, miR-26a, miR-23a, miR-200c, let-7a, and miR-1979 were identified as candidate reference genes for subsequent validation. MiR-23a was identified as the most reliable reference gene followed by miR-191. The use of miR-23a and miR-191 to normalize expression data enabled detection of a significant deregulation of miR-424 between normal, CIN and cancer tissue. Our results suggested that miR-23a and miR-191 are the optimal reference microRNAs that can be used for normalization in profiling studies of cervical tissues; miR-23a is a novel microRNA normalizer.

Evaluation of secondary cytoreduction surgery in platinum-resistant ovarian cancer patients within three-line recurrent: a multicenter, randomized controlled study

Ting-Ting Chen,Junfen Xu,Bairong Xia,Hui Wang,Yuanming Shen 대한부인종양학회 2024 Journal of Gynecologic Oncology Vol.35 No.1

Background: Epithelial ovarian cancer is the leading cause of death among gynecologicalmalignancies. Platinum resistance remains a dilemma and bottleneck in treatment, andsalvage chemotherapy has limited effectiveness. Recently, the role of secondar y cytoreductivesurger y (SCS) in patients with platinum-resistant recurrent ovarian cancer (ROC) has causedattention especially in patients with oligometastases. However, there is neither high-qualityevidence-based evidence nor standardized criteria for selecting SCS for patients withplatinum-resistant ROC until now. Methods: This multicenter, randomized, controlled clinical trial is to evaluate the valueof SCS and to clarif y reliable criteria of utilizing SCS in women with ROC, which is led byGynecologic Oncology Group, Women’s Hospital, Zhejiang University School of Medicine. Recruitment has started on Januar y 1st, 2023, and is scheduled to end in December 2026. One hundred and forty participants with platinum-resistant ROC who meet the “RSCScriteria” will be randomized assigned at a ratio of 1:1 to either the experimental arm or thestandard arm. Patients in the experimental arm will receive SCS followed by non-platinumsingle agent chemotherapy (paclitaxel, gemcitabine or liposomal adriamycin) for at least4 cycles while patients in the standard arm will be provided with only non-platinum singleagent chemotherapy. The primar y outcome is progression-free sur vival. The secondar youtcomes are overall sur vival, adverse events and health-related cancer-specific quality of life. Trial Registration: ClinicalTrials.gov Identifier: NCT05633199

Comparison of the safety between cervical conization and hysterectomy for patients with cervical adenocarcinoma in situ

Jingjing Liu,Yu Wang,Xiaoyun Wan,Jian Zou,Yedan Zhu,Weiguo Lv,Yuanming Shen 대한부인종양학회 2023 Journal of Gynecologic Oncology Vol.34 No.1

Objective: To compare the safety between cervical conization (CC) alone and hysterectomy for patients with adenocarcinoma in situ (AIS) of the cervix. Methods: Patients diagnosed with AIS after CC during 2007–2021 were identified by computerized databases at Women’s Hospital of Zhejiang University School of Medicine. A total of 453 AIS patients were divided into 2 groups according to uterus preservation: hysterectomy group (n=300) and CC(s) alone group (n=153). The prevalence of residual disease and disease recurrence was compared between patients treated by CC(s) alone and hysterectomy. The prevalence of residual disease in specimens from women who had a hysterectomy and repeat CC were compared between positive and negative margins of CC. The factors influencing residual disease and disease recurrence were assessed. Results: Among 310 specimens from women who had a hysterectomy or repeat CC, the prevalence of residual disease was 50.6% (45/89) for a positive margin and 2.3% (5/221) for a negative margin (p=0.000). Four patients had recurrence of vaginal intraepithelial neoplasia in those treated by hysterectomy and one had recurrence of cervical squamous intraepithelial neoplasia in those treated by CC(s) alone. The prevalence of recurrence was 0.7% (1/153) for CC(s) alone and 1.3% (4/300) for hysterectomy (p=0.431). Hysterectomy did not influence residual disease or disease recurrence. Conclusion: CC is an efficacious and safe option for patients with AIS of the cervix provided the margin is negative.

Efficacy of lymph node dissection on stage IIICr of cervical cancer before CCRT: study protocol for a phase III, randomized controlled clinical trial (CQGOG0103)

Misi He,Mingfang Guo,Qi Zhou,Ying Tang,Lin Zhong,Qing Liu,Xiaomei Fan,Xiwa Zhao,Xiang Zhang,Gang Chen,Yuanming Shen,Qin Xu,Xiao-jun Chen,Yuancheng Li,Dongling Zou 대한부인종양학회 2023 Journal of Gynecologic Oncology Vol.34 No.3

Background: Cervical cancer is still present a major public health problem, especially in developing countries. In International Federation of Gynaecology and Obstetrics 2018, allowing assessment of retroperitoneal lymph nodes by imaging and/or pathological findings and, if deemed metastatic, the case is designated as stage IIIC (with r and p notations). Patients with lymph node metastases have lower overall survival (OS), progression free survival (PFS), and survival after recurrence, especially those who have unresectable macroscopical positive lymph nodes. Retrospective analysis suggests that there may be a benefit to debulking macroscopic nodes that would be otherwise difficult to sterilize with standard doses of radiation therapy. However, there are no prospective study reporting that resecting macroscopic nodes before concurrent chemoradiation therapy (CCRT) would improve PFS or OS of cervical cancer and no guidelines for surgical resection of bulky lymph nodes. The CQGOG0103 study is a prospective, multicenter and randomized controlled trial (RCT) evaluating lymph node dissection on stage IIICr of cervical cancer. Methods: Eligible patients are histologically confirmed cervical squamous cell carcinoma, adenocarcinoma, adeno-squamous cell carcinoma. Stage IIICr (confirmed by computed tomography [CT]/magnetic resonance imaging/positron emission tomography/CT) and the short diameter of image-positive lymph node ≥15 mm. 452 patients will be equally randomized to receive either CCRT (pelvic external-beam radiotherapy [EBRT]/extended-field EBRT + cisplatin [40 mg/m2] or carboplatin [the area under curve=2] every week for 5 cycles + brachytherapy) or open/minimally invasive pelvic and para-aortic lymph node dissection followed by CCRT. Randomization is stratified by status of para-aortic lymph node. The primary endpoint is PFS. Secondary endpoints are OS and surgical complications. A total of 452 patients will be enrolled from multiple hospitals in China within 4 years and followed up for 5 years.