1.Long noncoding RNA HClnc1 promotes proliferation and migration of liver cancer cells by targeting RBBP5/KAT2B complex to enhance ODC1 transcription.
Zhihui FENG ; Wenyue LI ; Mingxiu ZHANG ; Peipei WANG ; Yangyang SHUAI ; Hong ZHANG
Journal of Southern Medical University 2025;45(9):1919-1926
OBJECTIVES:
To investigate the role of long noncoding RNA (lncRNA) HClnc1 in regulating proliferation, invasion, and migration of hepatocellular carcinoma (HCC) cells and the regulatory mechanism.
METHODS:
HClnc1 expression levels in liver cancer tissues were analyzed using data from the TCGA database. BrdU incorporation, plate cloning, and transwell assays were employed to examine the effects of HClnc1 silencing/overexpression and/or ODC1 silencing on proliferation, invasion, and migration of liver cancer cells. The effects of HClnc1 silencing on ODC1 protein and mRNA expression in the liver cancer cells were analyzed using qRT-PCR and Western blotting. The activity of ODC1 promoter was analyzed using a dual luciferase reporter gene assay. Pull-down experiment, mass spectrometry analysis, and chromatin immunoprecipitation (ChIP) assay were used for identification of HClnc1-binding proteins and their interactions. Protein interactions with the ODC1 promoter region and their binding efficiencies were investigated using RNA interference and ChIP analysis.
RESULTS:
HClnc1 was significantly overexpressed in HCC tissues. In liver cancer cells, HClnc1 silencing significantly inhibited cell proliferation, invasion, and migration, while HClnc1 overexpression promoted these behaviors. ODC1 silencing also suppressed malignant behaviors of liver cancer cells, and counteracted the effects of HClnc1 overexpression. Interference of HClnc1 obviously inhibited ODC1 promoter activity. RBBP5 and KAT2B proteins were identified to bind simultaneously with HClnc1. HClnc1 overexpression upregulated ODC1 protein expression, while interference of RBBP5 or KAT2B downregulated ODC1 protein expression and blocked HClnc1-induced upregulation of ODC1 protein. Both RBBP5 and KAT2B could directly bind to ODC1 promoter region; knocking out KAT2B or RBBP5 reduced the binding efficiency, while knocking out HClnc1 reduced the binding of both RBBP5 and KAT2B to ODC1 promoter region.
CONCLUSIONS
By targeting the RBBP5/KAT2B epigenetic modification complex, HClnc1 increases ODC1 promoter activity to enhance ODC1 transcription and promote the proliferation and migration of liver cancer cells.
Humans
;
Cell Proliferation
;
RNA, Long Noncoding/genetics*
;
Cell Movement
;
Liver Neoplasms/metabolism*
;
Cell Line, Tumor
;
Carcinoma, Hepatocellular/genetics*
;
Promoter Regions, Genetic
;
Gene Expression Regulation, Neoplastic
2.The value of multi-sequence MRI radiomics in predicting the early response to concurrent chemoradiotherapy for locally advanced cervical squamous cell carcinoma
Jing TONG ; Guiguang BEI ; Yangyang KAN ; Shuai HE ; Lijun WEI ; Yahong LUO
Chinese Journal of Radiology 2022;56(11):1223-1229
Objective:To investigate the predictive value of MRI radiomics model in assessing the early response to concurrent chemoradiotherapy for locally advanced cervical squamous cell carcinoma.Methods:A total of 367 patients with pathologically proven locally advanced cervical squamous cell carcinoma (International Federation of Gynecology and Obstetrics stage ⅡB-ⅣA) in Liaoning Cancer Hospital & Institute from January 2013 to June 2019 were retrospectively collected. The patients were unable to undergo surgery and received complete concurrent chemoradiotherapy. Pelvic plain MRI, DWI and dynamic contrast-enhanced MRI were performed within 2 weeks before treatment and at the end of the 4th week of treatment. Patients were divided into complete response (CR) group ( n=247) and non-CR group ( n=120) according to response evaluation criteria in solid tumors 1.1. The patients were divided into a training set ( n=256) and a validation set (n=111) via a randomized split method at a ratio of 7∶3. Two radiologists drew the region of interest on the DWI, T 2WI and contrast-enhanced T 1WI (delayed phase) images before treatment to form the volume of interest finally. Totally 1 906 radiomics features were extracted from 3 single sequence images, respectively. Feature correlation analysis and tree model were used for feature selection. Three classifier learning algorithms, namely logistic regression (LR), support vector machine and random forest, were used for machine learning and the best classifier was selected. Based on the best classifier, 3 single sequence radiomics models were built, and a multi-sequence combined model was obtained by multivariate LR analysis. The differences in the area under the receiver operating characteristic curve (AUC) of the 3 single sequence models and the multi-sequence combined model were compared by DeLong test. The clinical application value of the multi-sequence combined model was evaluated by decision analysis curve. Results:In the training set and validation set, the LR classifier model had the best performance. Based on the LR classifier, AUCs of DWI, T 2WI, contrast-enhanced T 1WI and combined sequences in the training set were 0.77, 0.74, 0.79 and 0.86, respectively, and AUCs in the validation set were 0.71, 0.66, 0.75 and 0.77, respectively. In the training set, the AUC of multi-sequence combined model was higher than those of DWI, T 2WI and contrast-enhanced T 1WI sequence models, and the differences were statistically significant ( Z=3.01, 3.56, 2.83; P=0.003, 0.001, 0.005). In the validation set, the AUC of multi-sequence combined model and T 2WI model had significant difference ( Z=2.46, P=0.015). The decision analysis curve showed that when the threshold probability was in the range of 0.44 to 0.88, the multi-sequence combined model yielded a net benefit. Conclusion:Based on the LR classifier, the combined model built by radiomics features of multi-sequence MRI images has predictive value for assessing the early response of concurrent chemoradiotherapy for locally advanced cervical squamous cell carcinoma.
3.Satisfaction degree and related factors of community health services among residents in Zhejiang province
Yangyang CHEN ; Shuai LI ; Jingjing REN ; Ying LIU
Chinese Journal of General Practitioners 2021;20(1):79-83
Objective:To analyze the satisfaction degree and related factors of community health services among residents in Zhejiang province.Methods:A service satisfaction survey was conducted during July to October 2019; using purpose sampling method three community health service centers were selected as survey sites in Hangzhou, Jiaxing, and Lishui cities, representing high, medium, and low economic development levels in Zhejiang province. The short-form Patient Satisfaction Questionnaire (PSQ-18) was used in the survey. SPSS 19.0 was adapted for data analysis, and multivariate logistic regression was applied use of analyze the affecting factors of satisfaction.Results:A total of 464 questionnaires were distributed, and 429 valid ones were returned with a response rate of 92.46%. Among them, 112 were from Hangzhou, 159 from Jiaxing, and 158 from Lishui. The overall satisfaction score of community health services was (4.22±0.59), and the satisfaction rate was 85.8%(368/429).Among the three cities, the overall satisfaction score in Lishui was the highest (4.28±0.51) with a satisfaction rate of 88.7% (140/158). The satisfaction scores of service quality, doctors' interpersonal communication ability, doctor-patient communication level, economic efficiency of service, timeliness of service, and accessability of service were (3.99±0.57), (4.10±0.77), (4.27±0.56), (3.54±0.84), (4.13±0.68) and (3.88±0.55); the satisfaction rates of those items were 59.6%(256/429), 79.4%(340/429), 86.3%(370/429), 43.6%(187/429), 78.9%(338/429)and 50.3%(216/429), respectively. Multivariate logistic regression analysis indicated that the quality of service, doctors' interpersonal communication ability, doctor-patient communication level and accessability of service were the affecting factors of the residents' satisfaction to community health services ( OR=3.734, 2.928, 7.732, 4.275; P<0.05). Conclusion:The residents of Zhejiang province have a high overall satisfaction with community health services. The quality of service, doctors' interpersonal communication ability, doctor-patient communication level and accessability of service are the affecting factors of the overall satisfaction degree.
4.Application of the saphenous nerve emerging site through the adductor canal in ultrasound-guided adductor canal block
Shuai TANG ; Xinhua SHEN ; Wei HUANG ; Manjiao MA ; Yangyang ZHANG ; Ying WANG ; Xu LI ; Xulei CUI ; Zhonghuang XU ; Chao MA ; Yuguang HUANG
The Journal of Clinical Anesthesiology 2018;34(2):114-117
Objective To study clinical data retrospectively and demonstrate the optimal injection site of adductor canal block by performing a cadaveric study.Methods Clinical part:clinical data from 19 patients,11 males and 8 females,aged 21 85 years,ASA physical status Ⅰ-Ⅲ,who received ultrasound guided adductor canal block were retrospectively collected.Among whom 9 received a mid-distance injection of 10 ml of 0.5% ropivacaine and 10 received an injection of the same medication at the outlet of adductor canal.The primary endpoint was complete absence of cold sensation to ice cube on the medial side of calf at 30 minutes and 24 hours after injection.Cadaveric part:40 lower limbs,20 males and 20 females,were finally analyzed in the study.The distances from the anterior superior iliac spine (ASIS) to the medial tibial condyle,from ASIS to the entrance of the adductor canal,from ASIS to the exit of the canal (adductor tendinous opening),from ASIS to the site where sa phenous nerve emerges through the aponeurotic covering were measured respectively.The length of adductor canal,the relative location of adductor canal and the site where saphenous nerve pierces in the lower limbs were calculated.Results Clinical part:all 19 cases were successfully recorded with complete absence of cold sensation at 30 minutes after injection of local anesthetic and complete sensory recovery at 24 hours after injection.Cadaveric part:in all specimens,saphenous nerve enters adductor canal and coursed down until emerging at very close to the distal end of the canal with the saphenous branch of descending genicular artery.The length of the adductor canal was (10.0±2.1) cm.The entrance and the exit of adductor canal and the emerging site of the saphenous nerve located along the (54.7±3.0) %,(76.0%±3.8) % and (74.1±3.2) % of sartorius muscle,respectively.Conclusion Performing ultrasound-guided adductor canal block at either the outlet of adductor canal or mid-distance of thigh can achieve comparable blockade of saphenous nerve.Cadaveric study implicated that the optimal injection site for adductor canal block should be the lower one-third of sartorius muscle.Ultrasound-guided injection of local anesthetics next to the descending genicular artery may possibly become a promising new method of saphenous nerve block.
5.Rapid cryopreservation for Siraitia grosvenorii cells based on cells' capacitance detection.
Jiarui LI ; Zejian WANG ; Meijin GUO ; Yuanxin GUO ; Shuai HUANG ; Yunfei SONG ; Zhen SUN ; Yangyang SUN ; Fanjing KONG ; Yingping ZHUANG
Chinese Journal of Biotechnology 2017;33(5):817-827
A rapid quantitative evaluation method for Siraitia grosvenorii cells was successfully developed based on plant cells' capacitance value detected by a viable cell mass monitor and the cryopreservation of S. grosvenorii suspension cells was optimized. The survival rate of S. grosvenorii cells was quantitatively measured by viable cell mass monitor and 2, 3, 5-triphenyltetrazolium chloride (TTC). An optimum cryoprotectant recipe is that the growth medium contained 10% sucrose and 10% DMSO. The experimental results also showed higher cell survival rates and cell viabilities were achieved when suspension cells were treated with pretreatment of 0.2 mol/L sucrose. With the increase of concentration of sucrose, however, the cell survival rate was decreased. And the cell survival rate represented a bell shape with the increase of pretreatment time. The highest cell survival rate and cell viability were obtained with the 9 h' s pretreatment. In addition, there was a good correlation between the cell survival rate measured by cell recovery test and that measured by viable cell mass monitor, while there were no significant differences in the cell morphology and the ability of mogrosides V production by S. grosvenorii cells cultured in suspension after cryopreservation. Therefore, the evaluation method developed based on the viable cell mass monitor has good feasibility and reliability.

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