1.Effects of miR-7-5p on the proliferation, apoptosis, and immune escape of esophageal squamous cell carcinoma KYSE-150 cells by regulating FOXM1
CHENG Lu1 ; LU Xiaoqun1 ; SUN Yan1 ; MEI Zhou1 ; WANG Jialu1 ; SUN Jie2
Chinese Journal of Cancer Biotherapy 2025;32(10):1044-1052
[摘 要] 目的:探讨miR-7-5p调控叉头框转录因子M1(FOXM1)对食管鳞状细胞癌(ESCC)KYSE-150细胞增殖、凋亡和免疫逃逸的影响。方法:通过双萤光素酶报告基因实验验证miR-7-5p与FOXM1的靶向结合位点。收集2022年1月至2024年10月期间苏州大学附属常州老年病医院收治的56例ESCC患者的癌组织和癌旁组织标本,以及患者的基本临床资料。采用qRT-PCR法检测ESCC组织中miR-7-5p和FOXM1的表达水平,分析其表达与临床病理特征的关系。常规培养的KYSE-150细胞为Ctrl组,用Lipofectamine 3000转染试剂将质粒转染KYSE-150细胞,分为Mimic NC组、miR-7-5p mimic组、miR-7-5p mimic + OE-NC组和miR-7-5p mimic + OE-FOXM1组。EdU染色和CCK-8实验检测KYSE-150细胞增殖能力,流式细胞术检测KYSE-150细胞的凋亡和CD8+ T细胞凋亡情况,WB法检测KYSE-150细胞中PD-L1、FOXM1、BAX和PCNA蛋白的表达。构建KYSE-150细胞裸鼠移植瘤模型,观察miR-7-5p过表达对移植瘤生长和组织中Ki-67、FOXM1表达的影响。结果:miR-7-5p可以靶向负调控FOXM1(P < 0.05)。ESCC组织中miR-7-5p呈低表达,FOXM1呈高表达(均P < 0.05),其表达分别与TNM分期、分化程度显著相关联(均P < 0.05)。miR-7-5p过表达组EdU阳性细胞率、细胞增殖能力、CD8+ T细胞凋亡率,以及PD-L1、PCNA、FOXM1 mRNA和蛋白均显著降低(均P < 0.05),细胞凋亡率、miR-7-5p、BAX水平均显著升高(均P < 0.05);同时过表达FOXM1则可逆转上述作用(均P < 0.05)。miR-7-5p过表达可降低小鼠移植瘤质量和体积,以及移植瘤组织中Ki-67、FOXM1蛋白的表达(均P < 0.05)。结论:miR-7-5p过表达能够显著抑制KYSE-150细胞增殖和免疫逃逸并促进细胞凋亡,其机制可能是通过靶向负调控FOXM1基因实现的。
2.Safety of allogeneic γδT cell immunotherapy for advanced hepatocellular carcinoma and its effect on patients' immune function
CHEN Yan1 ; ZHANG Yitian2, ; XU Yan3 ; LI Man3 ; LI Jiawei3 ; MENG Lingwen3 ; XIANG Zheng4 ; LIU Bing2 ; YIN Zhinan3 ; WU Bin1
Chinese Journal of Cancer Biotherapy 2024;31(3):253-260
[摘 要] 目的:探讨使用同种异体Vγ9Vδ2 T细胞回输治疗晚期肝细胞癌(HCC)患者的安全性及治疗后患者免疫功能的变化。方法:选择2021年10月至2022年10月珠海市人民医院收治的4例晚期HCC患者,从健康供体获取外周血单个核细胞(PBMC)后经刺激扩增培养获得Vγ9Vδ2 T细胞,经质控放行后予以回输治疗,回输细胞剂量为5×108个/次,每两周一次,回输次数9次以上,治疗后检测患者αβT细胞、B细胞、NK细胞、γδT细胞各亚群比例,转氨酶、肌酐、肌酸激酶等肝、肾、心功能生化标志物,以及血常规三系(白细胞系统、红细胞系统和血小板系统)细胞数量的变化。结果:4例患者在回输治疗后均显示出对异体Vγ9Vδ2 T细胞良好的耐受性;转氨酶、肌酐、肌酸激酶等肝、肾、心功能生化标志物以及血常规三系细胞数量在回输前后均无明显变化;患者的Tfh1、Tc1、CD127+TEM、HLADR+CD8+ T细胞、CD27- B细胞比例有升高趋势,提示特异性免疫功能的增强。结论:同种异体Vγ9Vδ2 T细胞治疗晚期HCC有较好的安全性并可在一定程度上改善患者的免疫功能。
3.Effects of overexpression of tyrosine kinase receptor 1 on malignant biological behavior of tumor-associated endothelial cells in cervical cancer cells
Qianyu1 Sun ; Yan1 Wei ; Rui1 Bai ; Ping1 Yang
Acta Universitatis Medicinalis Anhui 2022;57(11):1756-1762
Objective :
To investigate the effect of overexpression of tyrosine kinase receptor 1 (Tie-1) in cervical cancer cells on the malignant biological behavior of tumor-related endothelial cells (TRECs) .
Methods :
Immuno- histochemical method was used to detect the expression of Tie-1 in cervical cancer cells ( CCCs) and TRECs of 96 patients with cervical cancer,and to analyze the correlation between the expression of Tie-1 in TRECs and clinico- pathological features and prognosis of patients.HeLa cells overexpressing Tie-1 (Hela-Tie1OE) were constructed, and HeLa-Tie1OE was co-cultured with human umbilical vein endothelial cells ( HUVECs) by Transwell cell co- culture method to obtain cervical cancer TRECs.Western blot was used to analyze Tie-1 protein expression.The migration,invasion and tubulogenesis of TRECs were detected by cell scratch assay,Transwell invasion and migra- tion assay and tubulogenesis assay.
Results :
The expression of Tie-1 was positively correlated with CCCs and TRECs in 96 cervical cancer patients.The positive expression rate of Tie-1 in TRECs of patients with stage(FIGO) Ⅰ B2-ⅡA,tumor diameter ≥4 cm ,cervical muscle invasion depth ≥ 1 /2 full layer ,adenocarcinoma ,medium and low differentiation cervical cancer was higher than that of patients with Ⅰ A 1-Ⅰ B 1,tumor diameter<4 cm, cervical muscle invasion depth <1 /2 full layer,squamous carcinoma,and high differentiation cervical cancer,re- spectively.The differences were statistically significant (P<0. 05) .The expression of Tie-1 in TRECs of cervical cancer patients had no significant correlation with age,lymph node metastasis and lymphatic space invasion.The positive expression of Tie-1 in cervical cancer TRECs was negatively correlated with 5-year progression-free survival time and overall survival time (P<0. 05) .The Tie-1 expression of TRECs obtained by co-culture of Hela-Tie1OE and HUVECs was up-regulated ,and the invasion ,migration and tubulogenesis of TRECs cells were enhanced.
Conclusion
The high expression of Tie-1 in TRECs of cervical cancer is related to FIGO stage,tumor diameter, degree of differentiation,depth of cervical muscular invasion,type of cervical cancer,and poor prognosis of pa- tients.Overexpression of Tie-1 can promote TRECs invasion,migration and tubulogenesis.


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