1.Expression of TAGLN in colorectal cancer tissues and its effect on malignant biological behaviors of SW480 cells
CHEN Yu1 ; CUI Facai2 ; ZHANG Yinping1 ; ZHANG Bing1 ; WANG Yuanyuan1 ; XIA Qingxin1
Chinese Journal of Cancer Biotherapy 2021;28(1):67-73
[Abstract] Objective: To investigate the expression of transgelin (TAGLN) in colorectal cancer (CRC) tissues and its effect on the proliferation, migration and invasion of CRC SW480 cells. Methods: Surgically resected CRC tissues and corresponding para-cancerous tissues of 97 CRC patients from May 2015 to August 2016 in the Affiliated Tumor Hospital of Zhengzhou University were collected; In addition, CRC cell lines SW620, SW480, HCT116 and normal colorectal mucosal cell line FHC were also collected for this study. Immunohistochemical staining was used to detect the expression of TAGLN in CRC tissues, and the correlation between TAGLN and patients’ clinicopathological features was analyzed. Quantitative Real-time quantitative polymerase chain reaction (qPCR) and Western blotting (WB) were used to detect the mRNA and protein expressions of TAGLN in CRC cell lines. si-TAGLN and si-Ctrl were respectively transfected into SW480 cells by liposome transfection method. The effects of silencing TAGLN on the proliferation, migration and invasion of SW480 cells were detected by CCK-8, Wound-healing assay and Transwell assay, respectively; and the expression of EMT-related proteins E-cadherin, N-cadherin and vimentin were detected by WB. Results: The positive expression rate of TAGLN in CRC tissues was significantly higher than that in para-cancerous tissues (P<0.01), and TAGLN expression was correlated with TNM stage, degree of tumor differentiation and lymph node metastasis in CRC patients (P<0.05 or P<0.01). The mRNA and protein expression levels of TAGLN in SW480 cells were significantly higher than those in FHC cells (all P<0.01). After TAGLN silence, the proliferation, invasion and migration ability of SW480 cells were significantly reduced (all P<0.01), the expression level of E-cadherin in SW480 cells was increased, while the expression levels of N-cadherin and vimentin were decreased (all P<0.01). Conclusion: TAGLN is highly expressed in CRC tissues and cells. Silencing TAGLN can inhibit the proliferation, invasion and migration of CRC cells, suggesting that TAGLN plays an important role in the occurrence and development of CRC.
2.Observation on Choroidal Thickness of School-aged Children and Relevant Factors
Dan-yang1 WANG ; Qian2 TAN ; Ka Man3 CHUN ; Hui-yu1 GUO ; Cao1 JIAO ; Liang2 ZHANG ; Xing-wu1 ZHONG
Journal of Sun Yat-sen University(Medical Sciences) 2020;41(1):103-111
【Objective】To observe macular choroidal thickness and topographical variation in Chinese healthy and myopic children and to investigate the correlated factors. 【Methods】 A total of 196 myopic children treated at Hainan Province Eye Hospital were selected and divided into hyperopia ,emmetropia,low,moderate and severe myopia groups according to their spherical equivalents(SE). Axial length(AXL),subfoveal choroidal thickness(SFCT)and 1 mm ,3 mm nasal,temporal,superior,and inferior to the foveal choroidal thickness were recorded. Forty- five students from Tung Wah Group of Hospital′s MA Kam Chan Memorial Primary School in Hong Kong were also recruited in this 1-year longitudinal study. Children were grouped to myopic group and emmetropic or hyperopic group according to SE ,thickness changes of choroid were compared among children with or without myopic shift. The correlation among choroidal SE,and AXL variations were also investigated.【Results】Mean SFCT was 250(204~314)μm. The choroid was thinner at the nasal and inferior sectors ,the thinnest being at 3mm nasal to the fovea ,and the thickest at 1mm temporal to the fovea. Choroidal thickness in all orientations became thinner with progressively descending refractive degree. SFCT and 1 mm around fovea decreased most compared with other surrounding directions in all groups(Kruskal- Wallis test,P < 0.05). The choroidal thickness in subfovea and other sectors got thinned after 1 year follow-up time(Pair t test,P < 0.05).The variation of the choroidal thickness in fovea and surrounding positions was positively correlated with the change of SE and negatively related to the change of the AXL. The myopic group had a faster descent of SE and a faster growth of AXL than the emmetropic or hyperopic group. SFCT and surrounding choroidal thickness showed a progressive descent in the myopic group,but a slight decrease in the emmetropic or hyperopic groups.【Conclusions】There is significant topographic variation of choroidal at different regions of the macular. The choroidal thickness decreases faster in myopes. SFCT is more sensitive to myopic progression compared with surrounding regions.
3.Construction of a prognostic model with angiogenesis-related immune genes in ovarian cancer and analysis of tumor microenvironment
LYU Wei1 ; WANG Jiali1 ; LIU Tianxu1 ; WANG Yu1 ; LIU Lihua1,2,3
Chinese Journal of Cancer Biotherapy 2024;31(8):803-814
[摘 要] 目的:采用生物信息学方法探索卵巢癌中与血管生成相关的免疫基因(ARIG),探讨其与卵巢癌患者预后的关系,并说明不同预后患者肿瘤微环境和免疫治疗的潜在差异,为卵巢癌患者提供新的治疗靶点。方法:分别从TCGA和GEO数据库下载卵巢癌的转录组数据和生存数据。利用R软件分析差异表达基因,利用Pearson相关系数鉴定血管生成相关基因与免疫相关基因之间的相关性,筛选出差异表达的ARIG。通过Lasso回归分析构建预后模型,通过Cox分析临床特征和风险评分,将样本分为高风险组和低风险组。通过单样本基因集富集分析(ssGSEA)、肿瘤免疫功能障碍和排斥(TIDE)分析预后风险模型与免疫浸润、免疫治疗反应的相关性。最后,收集河北医科大学第四医院2015年5月至2016年5月手术的卵巢癌患者的肿瘤组织和输卵管组织85对,通过qPCR和WB法验证五个差异表达的ARIG在卵巢癌组织中的表达情况,分析其与卵巢癌患者临床病理特征的关系,并初步探索其在卵巢癌细胞的生物学功能。结果:通过生信分析筛选出142个差异表达的ARIG,通过Lasso和Cox回归分析,得到5个基因作为预后基因(PTGER3、SCTR、IGHG1、HSPA8、IGF2),构建了预后风险模型,高风险组患者的预后更差;此外,不同风险评分的患者在免疫细胞浸润和免疫治疗反应方面存在显著差异(均P<0.05)。通过qPCR和WB法验证这5个基因在卵巢癌组织中均为高表达(均P<0.01),其中HSPA8表达量最高,且高表达HSPA8与卵巢癌患者FIGO分期晚、组织分级差、淋巴结转移及腹膜转移呈显著正相关(P<0.001)。细胞功能实验证实,HSPA8可促进卵巢癌细胞的增殖、迁移和侵袭(P<0.01)。结论:差异表达的5种ARIG能有效预测卵巢癌患者的预后,并且与免疫细胞浸润和免疫治疗疗效有关,初步证实其在卵巢中发挥促癌作用。
4.The mechanism of BRCC3/NLRP3 in promoting the transformation of endometriosis to endometriosis-associated ovarian carcinoma
LIU Yu1 ; WU Qiongwei1 ; ZHANG Wenying1 ; WANG Chunchun1 ; HUANG Yuhua1 ; LI Bing1 ; MA Chengbin1 ; YANG Yu2
Chinese Journal of Cancer Biotherapy 2023;30(1):35-41
[摘 要] 目的:探讨NOD样受体蛋白3(NLRP3)炎症小体的活化在子宫内膜异位症(EMT)进展为EMT相关性卵巢癌(EAOC)过程中的作用及其机制。方法:选取2018年4月至2019年6月上海市长宁区幼保健院收治的EAOC、EMT、正常子宫内膜(CON组)组织标本各15例及患者的临床资料,利用免疫组织化学染色法、WB法检测EAOC、EMT和CON组织中NLRP3、caspase-1和IL-1β及含BRCA1/BRCA2的复杂亚基3(BRCC3)的表达水平。构建过表达BRCC3质粒和si-NLRP3质粒并转染EMT细胞CRL-7566,通过WB法检测转染后细胞中BRCC3蛋白的表达水平,利用MTT法、流式细胞术及Transwell实验分别检测转染后细胞增殖、凋亡、迁移与侵袭能力的变化。对过表达BRCC3组细胞进行干扰NLRP3实验,通过WB法检测干扰后BRCC3和NLRP3蛋白的表达水平,检测干扰后细胞增殖、凋亡、迁移与侵袭能力的变化。结果:EAOC和EMT组织中NLRP3、caspase-1、IL-1β和BRCC3的表达水平较CON组均呈明显升高(均P<0.01),且EAOC组织中NLRP3与BRCC3的表达呈正相关(r=0.65,P<0.01)。在CRL-7566细胞中过表达BRCC3显著促进细胞的增殖、迁移和侵袭并抑制细胞凋亡(均P<0.01),敲减NLRP3则抑制CRL-7566细胞的上述表型(均P<0.01),过表达BRCC3增强NLRP3的表达水平(P<0.01),而干扰BRCC3则抑制NLRP3表达(P<0.01);干扰NLRP3可以部分逆转BRCC3对细胞凋亡的抑制作用(P<0.01)、对细胞迁移(P<0.05)和侵袭(P<0.01)的促进作用。结论:EAOC和EMT组织中NLRP3和BRCC3均呈高表达,过表达BRCC3可促进CRL-7566细胞的增殖、迁移和侵袭并抑制细胞凋亡,与EMT向EAOC转化有关,BRCC3/NLRP3是潜在的EAOC炎癌转化预测标志物及治疗靶点。