目的:探讨新型重组杨梅素抑制4 T1细胞的作用及机制.方法:通过对杨梅素化学式改造合成一种新的杨梅素衍生物;CCK-8细胞增殖力抑制实验,并计算半数抑制浓度(IC50).划痕与Transwell实验观察新型杨梅素对4T1细胞迁移与侵袭能力的影响.流式细胞术观察4 T1细胞在合成杨梅素干预后其周期与凋亡的变化情况,小鼠体内移植瘤模型评估新型杨梅素在模拟体内环境下对小鼠三阴性乳腺癌(TNBC)的抑制作用.HE和TUNEL染色观察小鼠移植肿瘤组织切片的细胞坏死与凋亡.最后,Western blotting检测4T1细胞干预后其相关蛋白(p53、Bcl2、Bax、Caspase 3)的表达.结果:IC50为5.5μmol/mL.与DMSO组相比,5.5μmol/mL新型重组杨梅素在作用于4T1细胞24 h,细胞计数和迁移率均降低,差异有统计学意义(P<0.01).流式细胞术和肿瘤组织切片TUNEL染色结果显示,与DMSO组相比,5.5μmol/mL新型重组杨梅素细胞凋亡细胞数增高,G1期细胞比例较低,G2期细胞比率增高,肿瘤生长体积和肿瘤重量均较小,p53和Bcl2蛋白下调,Caspase 3上调,差异有统计学意义(P<0.05～P<0.01).结论:新型重组杨梅素可在体外和体内诱导4T1细胞凋亡,抑制其增殖.可能通过下调p53抑制4T1细胞增殖,通过Bcl2线粒体凋亡途径诱导4T1细胞凋亡.

目的 探讨杨梅素衍生物对人非小细胞肺癌A549细胞增殖及凋亡的影响和放射增敏作用.方法 将修饰后的杨梅素溶解在DMSO中,A549细胞被分成DMSO对照组和杨梅素衍生物组,MTT及Ki67实验检测A549细胞的增殖能力,Tran-swell检测A549细胞的侵袭能力,流式细胞术检测A549细胞的凋亡,Western blot法检测A549细胞中Bax、Bcl-2、P53、cleavedCaspase-3的表达情况.集落形成实验检测杨梅素衍生物对A549细胞的放射增敏作用.结果 与对照组比较,杨梅素衍生物组A549细胞活力降低(P<0.001),A549细胞的增殖指数下降(P<0.01),同时A549细胞的侵袭能力减弱(P<0.001),细胞凋亡比例增加(P<0.05).Western blot结果显示杨梅素衍生物处理后A549细胞中Bax/Bcl-2的比值增大,P53表达减少,cleaved Caspase-3表达增强(P<0.001).集落形成实验显示杨梅素衍生物的放射增敏参数SER为1.31.结论 杨梅素衍生物可通过诱导细胞凋亡相关蛋白的表达,降低A549细胞的增殖及侵袭能力,同时可作为放射增敏剂提高放疗疗效.

目的 对已发表的基于蛋白水平的恶性肿瘤放射耐受或抵抗相关研究进行定量分析,推荐出"放射耐受或抵抗领域值得进一步研究的蛋白".方法 检索词包括"RadiationTolerance"OR"radioresistance"AND"Neoplasms",在PubMed上检索2011-01-01-2020-06-30关于恶性肿瘤放射抵抗或耐受研究相关文献中所涉及蛋白水平的研究.梳理每个蛋白在所纳入文献中被研究的次数,定义为研究数量指数.以研究数量指数5次为界,分别设计高频组(≥5次)和低频组(<5次)的评价方案以进行推荐.高频组蛋白推荐方案基于研究质量指数评分表和贝叶斯统计法计算得到的累计排序概率图下面积值(SUCRA)进行.之后以研究数量指数和计算得到的研究质量指数均在中位数以上的蛋白作为高频组"放射耐受或抵抗领域值得进一步研究的蛋白"进行推荐.低频组蛋白采用R语言对<5与≥5次的放射耐受或抵抗相关蛋白分别进行信号通路富集后,以两者交集的信号通路中涉及的<5次的蛋白作为低频组"放疗领域值得进一步研究的蛋白"进行推荐.结果 截至2020-06-30,恶性肿瘤放射抵抗或耐受研究相关蛋白研究共1231篇,涉及蛋白共计830个.高频组蛋白51个,低频组779个.研究数量指数最高的为缺氧诱导因子-1A(HIF1A)蛋白,研究质量指数最高的为酪氨酸蛋白激酶(MET)蛋白.高频组所产生的"放疗领域值得进一步研究的蛋白"共9个,分别为NOTCH1、AKT1、EGFR、HSP90、PI3K、MMP2、RAD51、Survivin和DNA-PK.在低频组与高频组的蛋白富集所涉及信号通路中,交集有22条,低频组"放疗领域值得进一步研究的蛋白"为BRCC3、SIRT3和PLK4等58个蛋白.结论 在放射耐受或抵抗的既往研究相关蛋白中,基于评分表、贝叶斯排序法及R语言生信分析等定量研究方法可进一步研究的蛋白共计67种.但是,这些蛋白的临床确切价值尚待临床前瞻性研究样本验证.

Objective:To compare the survival rate and adverse reactions of patients with advanced hypopharyngeal squamous cell carcinoma undergoing surgery combined with chemoradiotherapy, and to analyze the prognostic factors of patients.Methods:The clinicopathologic data of 78 patients with advanced hypopharyngeal squamous cell carcinoma admitted to the Department of Radiation Oncology of the First Affiliated Hospital of Bengbu Medical University from August 2013 to December 2018 were retrospectively analyzed. The patients were divided into surgery combined with chemoradiotherapy group ( n=27) and chemoradiotherapy group ( n=51) according to different treatment methods. The median follow-up time was 46 months (20-84 months). The main observation indicators were overall survival (OS), progression-free survival (PFS) and local control rate (LCR). Cox regression model was used to analyze the prognostic factors. Results:Until July 31, 2020, 51 of the 78 patients with advanced hypopharyngeal squamous cell carcinoma died, including 6 cases of local recurrence, 11 cases of distant metastasis, and 34 cases of other causes (15 cases of hemorrhage, 15 cases of cachexia, and 4 cases of other diseases). In the surgery combined with chemoradiotherapy group, 12 patients died, accounting for 44.44%. In the chemoradiotherapy group, 39 patients died, accounting for 76.47%. The 1-, 3- and 5-year OS rates of 78 patients were 57.7%, 36.3% and 27.2% respectively, the 1-, 2- and 3-year PFS rates were 49.5%, 38.7% and 32.6% respectively, and the 1-, 2- and 3-year LCR were 53.4%, 40.0% and 34.2% respectively. The 1-, 3- and 5-year OS rates in the surgery combined with chemoradiotherapy group were 74.1%, 50.1% and 44.6%, and those in the chemoradiotherapy group were 49.0%, 29.3% and 12.8%, with a statistically significant difference ( χ2=5.142, P=0.023). The 1-, 2- and 3-year PFS rates in the surgery combined with chemoradiotherapy group were 62.1%, 54.3% and 44.4%, and those in the chemoradiotherapy group were 43.1%, 30.6% and 26.7%, with no statistically significant difference ( χ2=3.222, P=0.073). The 1-, 2- and 3-year LCR of the surgery combined with chemoradiotherapy group were 69.8%, 54.3% and 44.4%, and those in the chemoradiotherapy group were 45.1%, 32.9% and 29.6%, with no statistically significant difference ( χ2=3.576, P=0.059). The results of univariate analysis showed that tumor T stage ( χ2=7.140, P=0.008), N stage ( χ2=4.493, P=0.034) and treatment method ( χ2=5.142, P=0.023) were all independent influencing factors of the OS of patient with advanced hypopharyngeal squamous cell carcinoma; T stage ( χ2=5.807, P=0.016) and N stage ( χ2=6.587, P=0.010) were both independent influencing factors of PFS. The results of multivariate analysis showed that tumor T stage ( HR=2.121, 95% CI: 1.142-3.938, P=0.017), N stage ( HR=2.088, 95% CI: 1.144-3.811, P=0.016) and treatment method ( HR=0.430, 95% CI: 0.226-0.815, P=0.010) were all independent prognostic factors of the OS of patients with advanced hypopharyngeal squamous cell carcinoma; T stage ( HR=1.884, 95% CI: 1.011-3.510, P=0.046) and N stage ( HR=1.904, 95% CI: 1.058-3.429, P=0.032) were both independent prognostic factors of PFS. During the treatment period, there were statistically significant differences in the incidences of radioactive pharyngitis [7.41% (2/27) vs. 39.22% (20/51), χ2=8.821, P=0.003] and radioactive dermatitis [3.70% (1/27) vs. 29.41% (15/51), χ2=7.156, P=0.007] between the surgery combined with chemoradiotherapy group and the chemoradiotherapy group. However, there were no statistically significant differences in the incidences of radioactive oral mucositis [11.11% (3/27) vs. 17.65% (9/51), χ2=0.186, P=0.666], bone marrow suppression [37.04% (10/27) vs. 50.98% (26/51), χ2=1.381, P=0.240], pharynx infection [11.11% (3/27) vs. 5.88% (3/51), χ2=0.143, P=0.706] and tracheal fistula [7.41% (2/27) vs. 0 (0/51), P=0.117] between the two groups. Conclusion:The 1-, 3- and 5-year OS rates in the surgery combined with chemoradiotherapy group are higher than those in the chemoradiotherapy group, and the incidences of adverse reactions are low. T stage, N stage and treatment method are independent prognostic factors for OS of advanced hypopharyngeal squamous cell carcinoma patients, while T stage and N stage are independent prognostic factors for PFS.