Attenuated LKB1-SIK1 signaling promotes epithelial-mesenchymal transition and radioresistance of non–small cell lung cancer cells

Background: Radiotherapy is one of the main therapeutic approaches for non–small cell lung cancer(NSCLC). However, radioresistant cancer cells can eventually cause tumor relapse and even fatal metastasis. It is thought that radioresistance and metastasis could be potentially linked by epithelial?mesenchymal transition(EMT). In this study, we established radioresistant NSCLC cells to investigate the potential relationship among radioresistance, EMT, and enhanced metastatic potential and the underlying mechanism involving liver kinase B1(LKB1)?Salt?inducible kinase 1(SIK1) signaling.Methods: The radioresistant cell lines A549 R and H1299 R were generated by dose?gradient irradiation of the paren?tal A549 and H1299 cells. The radioresistance/sensitivity was evaluated by Cell Counting Kit?8 assay, apoptosis analysis, and/or clonogenic cell survival assay. The EMT phenotype and the signaling change were assessed by Western blot?ting. The abilities of invasion and migration were evaluated by transwell assays and wound healing assays.Results: The radioresistant cell lines A549 R and H1299 R displayed mesenchymal features with enhanced invasion and migration. Mechanistically, A549 R and H1299 R cells had attenuated LKB1?SIK1 signaling, which leaded to the up?regulation of Zinc?finger E?box?binding homeobox factor 1(ZEB1)—a transcription factor that drives EMT. Re?expression of LKB1 in A549 R cells reversed the EMT phenotype, whereas knockdown of LKB1 in H1299 R cells further promoted the EMT phenotype. Moreover, re?expression of LKB1 in A549 cells increased the radiosensitivity, whereas knockdown of LKB1 in H1299 cells decreased the radiosensitivity.Conclusions: Our findings suggest that attenuated LKB1?SIK1 signaling promotes EMT and radioresistance of NSCLC cells, which subsequently contributes to the enhanced metastatic potential. Targeting the LKB1?SIK1?ZEB1 pathway to suppress EMT might provide therapeutic benefits.

Liposome transfected to plasmid-encoding endostatin gene combined with radiotherapy inhibits liver cancer growth in nude mice

AIM: To evaluate whether intratumoral injection of liposomeendostatin complexes could enhance the antitumor efficacy of radiation therapy in human liver cardnoma (BEL7402) model. METHODS: Recombinant plasmid pcDNA3.End was transfected into human liver carcinoma cell line (BEL7402)with lipofectamine to produce conditioned medium. Then BEL7402 cells and human umbilical vein endothelial cells (HUVECs) were treated with the conditioned medium. Cell cycle and apoptosis were analyzed by flow cytometer and endothelial cell proliferation rates were determined by MTT assay. The antitumor efficacy of endostatin gene combined with ionizing radiation in mouse xenograft liver tumor was observed.RESULTS: Endostatin significantly suppressed the S phase fraction and increased the apoptotic index in HUVECs. In contrast, endostatin treatment had no effect on BEL7402 cell apoptosis (2.1±0.3% vs8.9±1.3%, t= 8.83, P= 0.009＜0.01)or cell cycle distribution (17.2±2.3% vs9.8±1.2%, t = 4.94,P = 0.016＜0.05). The MTT assay showed that endostatin significantly inhibited the proliferation of HUVECs by 46.4%.The combination of local endostatin gene therapy with radiation therapy significantly inhibited the growth of human liver carcinoma BEL7402 xenografts, the inhibition rate of tumor size was 69.8% on d 28 compared to the untreated group. The tumor volume in the pcDNA3.End combined with radiation therapy group (249±83 mm3) was significantly different from that in the untreated group (823±148 mm3, t= 5.86, P= 0.009＜0.01) or in the pcDNA3 group (717±94 mm3, t= 6.46, P= 0.003＜0.01). Endostatin or the radiation alone also inhibited the growth of liver tumor in vivo, but their inhibition effects were weaker than those of endostatin combined with radiation, the inhibition rates on d 28 were 44.7% and 40.1%, respectively.CONCLUSION: Endostatin not only significantly suppresses tumor growth but also enhances the antitumor efficacy of radiation therapy in human carcinoma xenograft.

Value of ^(18)F-FDG PET-CT in surveillance of postoperative colorectal cancer patients with various carcinoembryonic antigen concentrations

AIM:To evaluate the value of positron emission tomography(PET)/computerized tomography(CT)in surveillance of colorectal cancer(CRC)patients with different carcinoembryonic antigen(CEA)concentrations.METHODS:One hundred and six postoperative CRC patients who had suspected recurrence or metastasis and received fluorodeoxyglucose(FDG)PET/CT within one week were included in this study.The final diagnosis was confirmed by histological examination or clinicalfollow-up over at least six months.RESULTS:The sensitivity,specificity,and accuracy of FDG PET/CT were 95.2%,82.6%,and 92.5%,and94.8%,81.4%and 92.8%,respectively,in the caseand lesion-based analyses.The sensitivity and accuracy of FDG PET/CT significantly differed from CT in both analyses(χ2=8.186,P=0.004;χ2=6.201,P=0.013;χ2=13.445,P=0.000;χ2=11.194,P=0.001).In the lesion-based analysis,the sensitivity,specificity,and accuracy of FDG PET/CT in the abnormal CEA group were97.8%,82.6%,and 95.6%,compared with 81.3%,80%,and 80.6%for patients with normal CEA levels.In case-based analysis,the sensitivity,specificity,and accuracy of FDG PET/CT were 97.2%,77.8%,and 95%in abnormal CEA group.Only in lesion-based analysis,the sensitivity and accuracy of FDG PET/CT in the abnormal CEA group were significantly superior to those in the normal CEA group(χ2=6.432,P=0.011;χ2=7.837,P=0.005).FDG PET/CT changed the management in 45.8%of patients with positive scans.CONCLUSION:FDG PET/CT showed superior diagnostic value and is an advisable option in surveillance of postoperative CRC patients with a vague diagnosis.

Immunization of mice with concentrated liquor from male zooid of Antheraea pernyi

AIM: To study the effects of concentrated liquor from male zooid of Antheraea pernyi on immunological mice.METHODS: For each experiment, 40 mice were randomly divided into normal saline group (control group) and three tested groups that were administered different dosages of concentrated liquor from male zooid of A. pernyi and food for 15 d. The typical FSR and HC50 value, monocyte-phagocytic exponent Kand emendated monocyte-phagocytic exponent α were determined and calculated respectively.RESULTS: After 24 and 48 h, the FSR values of the three tested groups improved significantly in comparison to the control group by variance analysis. The HC50 values showed a significant difference between the high dosage group and the control group, as well as between the high dosage group and other two tested groups. The monocyte-phagocytic exponent Kand emendated exponent α showed rising tendencies, but no significant differences were found by variance analysis.CONCLUSION: The concentrated liquor from male zooid of A. pernyi can significantly enhance cellular and humoral immune function in mice, but has no distinct influence on the monocyte-phagocytic system in mice.

Ankle-brachial index as a predictor of all-cause and cardio- vascular disease mortality in 3733 Chinese patients with high cardiovascular risk

目的将在在高 CVD 风险的中国病人估计在所有原因和心血管的疾病(CVD ) 死亡和脚关节臂的索引(ABI ) 的 1 年风险之间的协会。完全，有高 CV 的 3733 个病人冒险的方法在基线有双边的 ABI 大小并且被跟随在上面为 1-1.5 年。病人们被划分到四个组：1 ) 冠的心疾病(CHD ) ；2 ) ischemic 击() ；3 ) 糖尿病 mellitus (DM ) ；4 ) 很高的风险组(VHR ) ，低 ABI 是被定义 < 0.9。3179 个病人全部的结果 A 被分析。低 ABI 的流行是 28.1% 。在 1 年，所有原因死亡是 8.7% ，和 27.6% 对 CVD 可归因；死亡由于 CV 事件是 4.8% 和 1.5% 。在调整另外的风险因素以后，低 ABI 的危险比率为所有原因死亡是 1.623 并且 2.304 为 CVD 死亡。在有或没有低 ABI 的病人类似，分别地在四 groups.Conclusion ABI 被发现是死亡的一个强壮、独立的预言者。有低 ABI 的病人有所有原因死亡和 CVD 死亡的实质地增加的风险。

Potential of Gd-EOB-DTPA as an imaging biomarker for liver injury estimation after radiation therapy

Background:Hepatic radiation injury severely restricts irradiation treatment for liver carcinoma.The purpose of this study was to investigate the clinical application of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)-enhanced MRI(EOB-MRI)in the assessment of liver function after external radiation therapy and to determine the relationship between focal liver reaction(FLR)and liver function.Methods:A total of 47 patients with liver malignancies who underwent external beam radiation therapy were enrolled.EOB-MRI was performed on each patient at approximately one month post-radiotherapy.The hepatobiliary(HPB)phase images from EOB-MRI were fused with the planning CT images,and the isodose lines from the patients’treatment plans were overlaid onto the fused images.The correlation of the EOB-MR image intensity distribution with the isodose lines was studied.We also compared liver function in patients between pre-treatment and post-treatment.Results:Decreased uptake of Gd-EOB-DTPA,which was manifested by well-demarcated focal hypointensity of the liver parenchyma or FLR to high-dose radiation,was observed in the irradiated areas of 38 patients.The radiotherapy isodose line of decreased uptake area of Gd-EOB-DTPA was 30–46 Gy.The median corresponding dose curve of FLR was 34.4 Gy.Nine patients showed the absence of decreased uptake area of Gd-EOB-DTPA in the irradiated areas.Compared to the 38 patients with the presence of decreased uptake area of Gd-EOB-DTPA,9 patients with the absence of decreased uptake area of Gd-EOB-DTPA showed significant higher levels of total bile acid,total bilirubin,direct bilirubin and alpha-fetoprotein(P<0.05).There were no significant differences in alanine transaminase,aspartate aminotransferase,gamma-glutamyl transpeptidase or albumin levels between the two groups(P>0.05).Conclusions:Visible uptake of Gd-EOB-DTPA by the liver parenchyma was significantly associated with liver function parameters.EOB-MRI can be a valuable imaging biomarker for the assessment of liver parenchyma function outside of radiation area.

Association between circulating CD39+CD8+T cells prechemoradiotherapy and prognosis in patients with nasopharyngeal carcinoma

Background:The mortality rate among patients with nasopharyngeal carcinoma(NPC)has improved significantly with the advent of chemoradiotherapy strategies.However,distant metastasis remains problematic.Tumor-specific reactivity in cancer patients has been detected exclusively in CD39+T cells,particularly in CD39+CD103+T cells.Circulating cancer-specific T cells are important for protecting against metastasis.This study aimed to evaluate the predictive value of circulating CD39+CD8+T cells for metastasis in patients with NPC.Methods:We performed a cross-sectional,longitudinal study of 55 patients with newly diagnosed NPC of stage III–IVa.All patients were initially treated with standard combined chemoradiotherapy.Blood samples were obtained from 24 patients before and at 1 month and 6 months after treatment.T cell expression of CD39 and CD103,together with the markers of T cell exhaustion programmed death-1(PD-1)/T cell immunoglobulin and mucin domain-containing protein 3(Tim-3)and markers of cell differentiation CD27/CC-chemokine receptor 7/CD45RA,was examined by flow cytometry.The Wilcoxon rank-sum test analysis was used to analyze the differences between two groups.Kaplan-Meier analysis was used for analysis of progression-free survival(PFS).Results:The expression of circulating CD39+CD8+and CD39+CD103+CD8+T cells was significantly higher in patients without distant metastasis(CD39+CD8+:6.52%[1.24%,12.58%]vs.2.41%[0.58%,5.31%],Z=2.073,P=0.038 and CD39+CD103+CD8+:0.72%[0.26%,2.05%]vs.0.26%[0.12%,0.64%],Z=2.313,P=0.021).Most CD39+T cells did not express PD-1 or Tim-3.Patients with high expression of CD39+CD103+CD8+T cells had better PFS than patients with low expression(log rank value=4.854,P=0.028).CD39+CD8+T cells were significantly elevated at 1-month post-treatment(10.02%[0.98%,17.42%]vs.5.91%[0.61%,10.23%],Z=2.943,P=0.003).The percentage of advanced differentiated CD8+T cells also increased at 1-month post-treatment compared with pre-treatment(33.10%[21.60%,43.05%]vs.21.00%[11.65%,43.00%],Z=2.155,P=0.031).There was a significant correlation between elevated CD39+CD8+T cells and increased effector memory T cells(intermediate stage:r=0.469,P=0.031;advanced stage:r=0.508,P=0.019).Conclusions:CD39+CD8+circulating T cells have preserved effector function,contributing to an improved prognosis and a reduced risk of metastasis among NPC patients.These cells may thus be a useful predictive marker for a better prognosis in patients with NPC.