Aim: To observe the efficacy of the low dose apatinib plus deep hyperthermia as third-line or later treatment for patients with human epidermal growth factor receptor 2 (HER-2) negative advanced gastric cancer. Methods: 80 eligible patients with HER-2 negative advanced gastric cancer admitted to Jingjiang People’s Hospital Affiliated with Yangzhou University-from March 2021 to March 2022 were selected, and they were divided into the control group ( n = 40, apatinib) and experimental group ( n = 40, apatinib plus deep hyperthermia) on the basis of random number table method. The levels of serum carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA199), and vascular endothelial growth factor (VEGF) were monitored, and the efficacy of the two groups was analyzed by referring to Karnofsky performance status (KPS), overall survival (OS) and disease control rate (DCR) before and after treatment. Results: The levels of CEA, CA199, and VEGF in both groups were lower after treatment than before ( p < 0.05), and lower (CEA: 8.85 ± 1.36 vs. 12.87 ± 1.23, CA199: 34.19 ± 4.68 vs. 50.11 ± 5.73, VEGF: 124.8 ± 18.03 vs. 205.9 ± 19.91) in the experimental group than in the control group ( p < 0.05). The DCR and KPS of the patients in the experimental group were significantly higher (DCR: 62.50% vs. 40.00%; KPS: 83.25 ± 1.15 vs. 76.25 ± 1.17) than in the control group ( p < 0.05). In survival analysis, patients with control group had shorter OS than the experimental group. (median 5.65 vs. 6.50 months; hazard ratio [HR], 1.63 [95% confidence interval (CI) 1.02–2.60], p = 0.0396). Conclusion: The application of low-dose apatinib plus deep hyperthermia for patients with HER-2 negative gastric cancer who failed second-line treatment should be a promising option.

目的 评估低剂量阿帕替尼联合热疗应用于二线治疗失败后的晚期胃癌患者的临床疗效和安全性.方法 随机选取2020年12月—2021年12月扬州大学附属靖江市人民医院收治的80例二线治疗失败的晚期胃癌患者为研究对象,根据随机数表法分为联合组和靶向治疗组,各40例,靶向治疗组患者口服阿帕替尼250 mg/d治疗,联合组患者在对照组基础上联合热疗,观察两组患者的临床疗效、不良反应、KPS及癌痛评分.结果 联合组DCR(90.0％)优于靶向治疗组(72.5％),差异有统计学意义(χ2=4.021,P=0.045).联合组与靶向治疗组的不良反应发生率比较,差异无统计学意义(P>0.05).联合组KPS评分好转率(65.0％)优于靶向治疗组(37.5％),差异有统计学意义(χ2=6.054,P=0.014).联合组癌痛评分低于靶向治疗组,差异有统计学意义(P<0.001).结论 对于晚期的胃癌患者采用口服低剂量阿帕替尼联合热疗具有较好的临床治疗效果,且不良反应发生率低,能够确保抗肿瘤治疗的有效性和安全性,可作为临床晚期胃癌患者的一种治疗选择.

BackgroundThis study aimed to explore the clinical efficacy and safety of a modified FOLFOX6 (oxaliplatin + leucovorin + 5-fluorouracil) plus bevacizumab regimen after deep hyperthermia in advanced colorectal cancer.MethodsA total of 80 colorectal cancer patients treated at our hospital were selected as research subjects. According to the random number table method, patients were divided into a control group (mFOLFOX6 plus bevacizumab) and a combination group (mFOLFOX6 plus bevacizumab after deep hyperthermia treatment), with 40 patients in each group. After six cycles of treatment, the objective response rate (ORR), disease control rate (DCR), levels of serum tumor markers carcinoembryonic antigen (CEA), vascular epidermal growth factor (VEGF), Karnofsky performance status (KPS) scores, and the occurrence of adverse events were compared between the two groups.ResultsAfter six cycles of treatment, the ORR in the combination group was higher than that in the control group, but the difference was not statistically significant (P>0.05). The DCR in the combination group was significantly higher than that in the control group (P<0.05). The serum CEA levels in the control and combination groups after treatment were significantly lower than those before treatment, and the serum CEA and VEGF levels in the combination group were significantly lower than those in the control group (all P<0.001). The KPS scores in both groups after treatment were higher than those before treatment, and the KPS scores in the combination group after treatment were significantly higher than those in the control group (all P<0.001). The incidence of fatigue and pain in the combination group was significantly lower than that in the control group (P<0.05).ConclusionmFOLFOX6 plus bevacizumab after deep hyperthermia is effective in advanced colorectal cancer patients, which can effectively improve their quality of life, and the adverse events are controllable and tolerable. A randomized or prospective trial will be required to further prove these data and explore its potentiality, especially if compared to conventional treatment.

目的 探讨深部热疗辅助安罗替尼治疗晚期肺腺癌患者的临床疗效和安全性.方法 筛选2020年6月至2021年12月扬州大学附属靖江市人民医院肿瘤内科收治的46例晚期肺腺癌患者,根据随机数字表法分为对照组(安罗替尼组)和治疗组(安罗替尼联合深部热疗组),每组患者各23例.治疗12周后进行影像学和相关血液学评估.主要观察晚期肺腺癌患者的临床疗效、症状改善、不良反应发生率、血清肿瘤标志物和生活质量等指标.结果 治疗组患者客观缓解率(objective response rate,ORR)(65.21％vs.34.78％)和疾病控制率(disease control rate,DCR)(91.30％vs.60.87％)均优于对照组,差异有统计学意义(χ2=4.261,5.855;P=0.039,0.016).治疗组患者咳嗽、气急、胸痛、疲乏、咳血、食欲下降好转率均优于对照组,差异有统计学意义(P<0.05).治疗期间,治疗组患者胃肠道反应发生率低于对照组,差异有统计学意义(P<0.05).两组患者治疗前的癌胚抗原(carcinoembryonic antigen,CEA)、血管内皮生长因子(vascular endothelial growth factor,VEGF)水平及其阳性率均高于治疗后;治疗后对照组患者的CEA、VEGF水平及其阳性率均高于治疗组,差异均具有统计学意义(P<0.05).治疗后治疗组患者的CD3、CD4水平均高于治疗前(P<0.05),治疗组患者的CD3、CD4水平均高于对照组;两组患者治疗前的CD3、CD4水平均低于治疗后,差异均具有统计学意义(P<0.05).治疗后治疗组患者生活质量评分优于对照组(65.22％vs.34.78％),差异有统计学意义(χ2=4.261,P=0.039).结论 深部热疗辅助安罗替尼治疗晚期肺腺癌患者可明显提高ORR和DCR,缓解患者咳嗽、气急、胸痛、疲乏、咳血、食欲下降等不适表现,有效减轻药物不良反应,提升患者免疫功能,改善生活质量.

Abstract The authors have requested that this preprint be removed from Research Square.

Radiotherapy is an effective therapeutic strategy in esophageal squamous cell carcinoma (ESCC). However, acquired radioresistance of cancer cells leads to radiotherapy failure. The present study aimed to investigate the mechanisms of the effect of high mobility group box 1 (HMGB1) on the radiation sensitivity of ESCC. Small interfering RNA (si) transfection was used to generate three groups of TE-1 cells (TE-1, negative control and TE-1+siHMGB1), and the protein expression levels of HMGB1 in TE-1 cells were detected by western blotting. These groups of TE-1 cells were irradiated with different doses (0, 2, 4, 6 and 8 Gy) of X-rays after transfection. Subsequently, the viability of TE-1 cells was detected using an MTT assay, and the survival fraction of TE-1 cells was observed using a colony formation assay. The apoptotic rate, reactive oxygen species (ROS) content and levels of phosphorylated (p)-histone H2AX at S139 (p-γH2AX) of the cells were detected by flow cytometry. The alterations in mRNA expression levels of nicotinamide adenine nucleotide phosphate oxidase (NOX)1 and NOX5 were detected by reverse transcription-quantitative PCR, while the changes in protein levels of caspase-3, poly(ADP-ribose) polymerase, p-p38, p-ERK1/2 and p-JNK were detected by western blotting. The results revealed that HMGB1 knockdown significantly decreased cell viability, and the apoptosis rate of TE-1 cells transfected with siHMGB1 combined with radiation treatment was increased compared with that in cells with either siHMGB1 transfection or radiation treatment alone. HMGB1 knockdown increased nicotinamide adenine nucleotide phosphate oxidase-mediated ROS production and induced DNA damage via the MAPK signaling pathway, which may promote apoptosis and radiosensitivity after radiation in TE-1 cells. In conclusion, targeting HMGB1 may represent a promising strategy to increase the efficacy of radiation therapy for ESCC.

目的 观察表没食子儿茶素没食子酸酯(EGCG)联合放射治疗(放疗)对食管癌TE-1细胞凋亡的影响,并探讨其分子机制.方法 选择人食管癌细胞株TE-1作为研究对象,并分成对照组、EGCG组、放疗组和联合组(EGCG+放疗).MTT法检测不同浓度EGCG、不同放射剂量单用和联用对食管癌TE-1细胞生长的影响;流式细胞术检测不同处理组细胞凋亡情况;Western blotting检测经不同处理后TE-1细胞凋亡相关蛋白cleaved caspase-3和抗凋亡蛋白Bcl-2的表达情况.结果 EGCG和放疗均能抑制食管癌TE-1细胞的增殖,但EGCG与放疗联合的抑制作用显著高于EGCG组或放疗组(P＜0.01),呈浓度-剂量依赖性.流式细胞术检测结果显示,与单独放疗组细胞相比,EGCG联合放疗可以明显提高X射线诱导的TE-1细胞凋亡(P＜0.01),上调cleaved caspase-3的表达(P＜0.05)和下调Bcl-2蛋白的表达(P＜0.05).结论 EGCG能够协同放疗增加食管癌TE-1细胞的凋亡,这种作用可能与cleaved caspase-3和Bcl-2有关.

目的:探讨外源性IL-23通过PI3 K/AKT/c-Myc通路对TE-1细胞上皮-间质转化(epithelial-mesenchymaltransition,EMT)和迁移的影响.方法:采用免疫组织化学和免疫荧光检测12例食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)患者癌组织与配对癌旁组织中IL-23及其受体(IL-23 R)的表达和胞内定位;蛋白质印迹检测TE-1细胞与阴性对照NIH3T3细胞中IL-23p19表达,流式细胞术检测IL-23R的表达;PCR和蛋白质印迹法检测空白对照组,50 ng/mL IL-23组,1μmol/L Wortmannin+50 ng/mL IL-23组细胞中c-Myc mRNA及其蛋白与E-钙黏蛋白、波形蛋白、p-AKT、Snail 1、Slug蛋白的表达;划痕和Transwell实验检测各组TE-1细胞侵袭和迁移能力.结果:与癌旁组织比较,ESCC组织及其细胞质中IL-23呈高表达,细胞膜上IL-23R高表达;与阴性对照比较,TE-1细胞中IL-23p19及IL-23R呈过表达(P均＜0.01);与对照组比较,IL-23组细胞中c-Myc mRNA及其蛋白、p-AKT、波形蛋白、Snail 1、Slug蛋白明显升高,E-钙黏蛋白表达明显降低,TE-1细胞划痕愈合率和迁移数明显升高(P均＜0.05).与IL-23组比较,IL-23+ Wortmannin组细胞中c-Myc mRNA及其蛋白、p-AKT、波形蛋白、Snail 1、Slug蛋白明显降低,E-钙黏蛋白明显升高,TE-1细胞划痕愈合率和迁移数明显降低(P均＜0.05).结论:IL-23通过PI3K/AKT/c-Myc通路促进食管鳞癌细胞上皮间质转化和迁移.

The PI3K/Akt/mTOR pathway is activated in a variety of human tumors including B-cell non-Hodgkin lymphoma (B-NHL). Targeting this pathway has been validated in solid and hematological tumors. In the present study, we demonstrated that PF-04691502, a novel PI3K/mTOR inhibitor has potent activity in a panel of aggressive B-NHL cell lines including diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL). MTS analysis showed that PF-04691502 effectively inhibited cell proliferation with IC50 values ranging from 0.12 to 0.55 µM. Cells treated with PF-04691502 exhibited decreased phosphorylation of Akt and S6 ribosomal protein confirming the mechanism of action of a PI3K/mTOR inhibitor. Also, treatment of B-NHL cell lines with PF-04691502 induced apoptosis in a dose- and time-dependent manner. Moreover, PF-04691502 significantly induced G1 cell cycle arrest associated with a decrease in cyclin D1 which contributed to suppression of cell proliferation. Finally, rituximab enhanced apoptosis induced by PF-04691502. Taken together, our findings provide for the first time that PF-04691502 inhibits the constitutively activated PI3K/mTOR pathway in aggressive B-cell NHL cell lines associated with inhibition of cell cycle progression, cell proliferation and promotion of apoptosis. These findings suggest that PF-04691502 is a novel therapeutic strategy in aggressive B-cell NHL and warrants early phase clinical trial evaluation with and without rituximab.

Objective:To detect the progress of epithelial-mesenchymal transition(EMT) in esophageal squamous carcinoma cells(ESCC) TE-1 which was treated with ionizing radiation,and explore the underlying mechanisms.Methods:After treated with different intensity of ionizing radiation (0,2,4,8 Gy),realtime PCR was used to detect the mRNA expression of Snail 1 and AP-1 in TE-1 cells.Western blot was used to measure the protein expressions of Snail 1,Slug,E-cadherin and Vimentin.The protein levels of Ecadherin and Vimentin was verified by immunofluorescence.The ability of migration was measured by wound healing test and Transwell inserts.Dealing with ionizing radiation (4 Gy) and/or DAPT the inhibitor of Notch signaling pathway,the expression levels of Notch 1,NICD,Hes 1 and MMP-9 proteins were measured by Western blot.Results:After treated with ionizing radiation,TE-1 cells showed the up-regulation effect of the transcription factors of EMT(Snail 1,Slug,AP-1),mesenchymal marker(Vimentin) and metastasis related protein (MMP-9),meanwhile the down-regulation effect of epithelial marker(E-cadherin).and when dealing with 4 Gy radiation,the changes was significant (P ＜0.01).Ionizing radiation accelerated the transfer ability of TE-1 (P ＜ 0.05).By up-regulating the expression levels of Notch 1/NICD/Hes 1 signaling pathway,ionizing radiation promoted EMT related protein expressions (P ＜ 0.01).Conclusion:The ionizing radiation strengthens EMT,thereby induces metastasis,by activating Notch signaling pathway.

As the eighth most common malignant tumour worldwide, oesophageal cancer (OC) is often diagnosed during the metastasis of its advanced stage. Interleukin (IL)-23 is an immunomodulatory cytokine that has recently been identified as a cancer-associated factor. However, the role of IL-23 in the evolution of OC remains unclear. In the present study, we found that IL-23 was significantly expressed in the tumours of OC patients suffering metastasis and demonstrated that IL-23 contributed to epithelial-mesenchymal transition (EMT) through the Wnt/β-catenin pathway, promoting the migration and invasion of OC cells. In conclusion, IL-23 plays a pivotal role in the development of OC via EMT.