Neoantigen cancer vaccines:a new star on the horizon

Immunotherapy represents a promising strategy for cancer treatment that utilizes immune cells or drugs to activate the patient's own immune system and eliminate cancer cells.One of the most exciting advances within this field is the targeting of neoantigens,which are peptides derived from non-synonymous somatic mutations that are found exclusively within cancer cells and absent in normal cells.Although neoantigen-based therapeutic vaccines have not received approval for standard cancer treatment,early clinical trials have yielded encouraging outcomes as standalone monotherapy or when combined with checkpoint inhibitors.Progress made in high-throughput sequencing and bioinformatics have greatly facilitated the precise and efficient identification of neoantigens.Consequently,personalized neoantigen-based vaccines tailored to each patient have been developed that are capable of eliciting a robust and long-lasting immune response which effectively eliminates tumors and prevents recurrences.This review provides a concise overview consolidating the latest clinical advances in neoantigen-based therapeutic vaccines,and also discusses challenges and future perspectives for this innovative approach,particularly emphasizing the potential of neoantigen-based therapeutic vaccines to enhance clinical efficacy against advanced solid tumors.

Autophagy inhibition enhances apigenin-induced apoptosis in human breast cancer cells

Apigenin （4＇,5,7-trihydroxyflavone） is a member of the flavone subclass of flavonoids present in fruits and vegetables. The involvement of autophagy in the apigenin-induced apoptotic death of human breast cancer cells was investigated. Cell proliferation and viability were assessed by 3-（4,5-dimethylthiazol- 2-yl）-2,5-diphenyltetrazolium bromide （MTT） and clonogenic assays. Flow cytometry, fluorescent staining and Western blot analysis were employed to detect apoptosis and autophagy, and the role of autophagy was assessed using autophagy inhibitors. Apigenin dose- and time-dependently repressed the proliferation and clonogenic survival of the human breast cancer T47D and MDA-MB-231 cell lines. The death of T47D and MDA-MB-231 cells was due to apoptosis associated with increased levels of Caspase3, PARP cleavage and Bax/Bcl-2 ratios. The results from flow cytometry and fluorescent staining also verified the occurrence of apoptosis. In addition, the apigenin-treated cells exhibited autophagy, as characterized by the appearance of autophagosomes under fluorescence microscopy and the accumulation of acidic vesicular organelles （AVOs） by flow cytometry. Furthermore, the results of the Western blot analysis revealed that the level of LC3-Ⅱ, the processed form of LC3-Ⅰ, was increased. Treatment with the autophagy inhibitor, 3-methyladenine （3-MA）, significantly enhanced the apoptosis induced by apigenin, which was accompanied by an increase in the level of PARP cleavage. Similar results were also confirmed by flow cytometry and fluorescence microscopy. These results indicate that apigenin has apoptosis- and autophagy-inducing effects in breast cancer cells. Autophagy plays a cyto-protective role in apigenin-induced apoptosis, and the combination of apigenin and an autophagy inhibitor may be a promising strategy for breast cancer control.

Interpretation of breast cancer screening guideline for Chinese women

Breast cancer is the most common malignant tumor in Chinese women.Early screening is the best way to improve the rates of early diagnosis and survival of breast cancer patients.The peak onset age for breast cancer in Chinese women is considerably younger than those in European and American women.It is imperative to develop breast cancer screening guideline that is suitable for Chinese women.By summarizing the current evidence on breast cancer screening in Chinese women,and referring to the latest guidelines and consensus on breast cancer screening in Europe,the United States,and East Asia,the China Anti-Cancer Association and National Clinical Research Center for Cancer(Tianjin Medical University Cancer Institute and Hospital)have formulated population-based guideline for breast cancer screening in Chinese women.The guideline provides recommendations on breast cancer screening for Chinese women at average or high risk of breast cancer according to the following three aspects:age of screening,screening methods,and screening interval.This article provides more detailed information to support the recommendations in this guideline and to provide more direction for current breast cancer screening practices in China.

Epigenetic modulation of the tumor immune microenvironment by nanoinducers to potentiate cancer immunotherapy

Epigenetic dysregulation is a key factor leading to oncogenesis and tumor progression1. To date, seven agents in three epigenetic target classes have been approved by the U.S. Food and Drug Administration for the treatment of diverse malignancies. Despite this approval for clinical application, the mechanisms of many epigenetic modulators are not completely understood, because many genes are epigenetically regulated.

The mutation landscape of multiple cancer predisposition genes in Chinese familial/hereditary breast cancer families

Objective:Approximately 5%–10%of breast cancer(BC)patients display familial traits that are genetically inherited among the members of a family.The purpose of this study was to profile the germline mutations in 43 genes with different penetration rates and their correlations with phenotypic traits in Chinese familial BC families.Methods:Ion Torrent S5™-based next generation sequencing was conducted on 116 subjects from 27 Chinese familial BC families.Results:Eighty-one germline mutations in 27 BC predisposition genes were identified in 82.8%(96/116)of the cases.Among these,80.8%of the mutated genes were related to DNA damage repair.Fourteen possible disease-causing variants were identified in 13 of 27 BC families.Only 25.9%(7/27)of the BC families exhibited hereditary deficiency in BRCA1/2 genes,while 22.2%of the BC families exhibited defects in non-BRCA genes.In all,41.7%(40/96)of the mutation carriers had BRCA mutations,88.5%(85/96)had non-BRCA mutations,and 30.2%(29/96)had both BRCA and non-BRCA mutations.The BC patients with BRCA mutations had a higher risk of axillary lymph node metastases than those without mutations(P<0.05).However,the BC patients with non-BRCA mutations frequently had a higher occurrence of benign breast diseases than those without mutations(P<0.05).Conclusions:In addition to BRCA1/2,genetic variants in non-BRCA DNA repair genes might play significant roles in the development of familial/hereditary BC.Therefore,profiling of multiple BC predisposition genes should be more valuable for screening potential pathogenic germline mutations in Chinese familial/hereditary BC.

Analysis of Lymph Node Metastases of 1,526 Cases with Thoracic Esophageal and Cardiac Carcinomas: A Random Sampling Report froni the Fourth Hospital of Hebei Medical University from 1996 to 2004

OBJECTIVE To summarize the regular pattern and state oflymph node metastasis of patients with esophageal and cardiaccarcinomas,so as to analyze factors influencing lymph nodemetastasis.METHODS Clinical data collected from 1,526 thoracicesophageal and cardiac carcinoma patients who were admitted inthe Fourth Hospital of Hebei Medical University during a periodfrom January 1996 to December 2004,were randomly selectedand an Access Database of the patient's information was set up.Eight clinico-pathologic factors,including the patient's age,tumorlocation and size,pathological classification,the depth of tumorinvasion,vascular tumor embolus (VTE),the state of surroundingorgan encroachment and the status of tumor residues,wereidentified.A correlation between these factors and metastases wasstatistically analyzed using SPSS13.0 software.RESULTS Lymph node metastatic sites from esophagealcarcinomas included the thoracic and abdominal cavity.Lymphnode metastasis from the superior esophageal carcinomasmainly occurred in the neck and thoracic cavity.There was atwo-way lymph node metastasis in the patients with the middleesophageal carcinoma.The inferior esophageal carcinomas mainlymetastasized to the paraesophageal,paragastric cardia,and leftgastric artery lymph nodes.The rate and degree of the metastasisfrom the inferior esophageal carcinomas were significantly highercompared to those of the superior and the middle esophagealcarcinomas (P<0.0125).The degree of abdominal lymph node metastasis fromcarcinomas of the gastric cardia was significantly higher comparedwith that of esophageal carcinomas.In the group with carcinomaof the gastric cardia,the rate and degree of the lymph nodemetastases in the paragastric cardia and left gastric artery weresignificantly higher compared to the group with esophagealcarcinoma (P<0.05).Paraesophageal lymph node metastasis fromcarcinomas of the gastric cardia in the thoracic cavity frequentlyoccurred,too,and the degree of the metastasis was similar to thatof esophageal carcinoma.There was no significant difference inthe rate and degree of the paraesophageal lymph-node metastasisbetween the group with carcinoma of the gastric cardia comparedto those with esophageal carcinoma (P>0.05).Multifactoriallogistic regression analysis showed that the tumor size,depth oftumor encroachment,VTE,and tumor residues could all bringabout obvious impact on lymph-node metastases (P<0.05).CONCLUSION Lymph node metastasis from superioresophageal carcinomas mainly occurs in the neck and thoraciccavity.The middle esophageal carcinomas presented a two-waylymph-node metastasis (both the upwards and the downwards),and the lymph node metastasis from inferior esophagealcarcinomas mainly occurred in the thoracic and abdominal cavities.The metastases of carcinoma of the gastriccardia were most commonly found in the abdominalcavity,with frequent paraesophageal lymph-nodemetastasis.The sufficient attention should be paidto neck lymph node clearance in cases of esophagealcarcinoma.What is of the greatest concern is theclearance of the left gastric artery lymph nodes,andalso in cases of gastric cardia carcinoma,clearance,the paraesophageal lymph nodes.With an increasein the tumor size and depth of tumor encroachment,and occurrence of VTE and tumor residual cells,therisk of lymph node metastasis is significantly raised (P<0.05).

Characterization of Natural Killer Cells in the Liver of Young Mice

OBJECTIVE To determine the quantity and quality of liver NK cells from young and adult mice and compare their characteristics. METHODS C57BL/6 mice were used at 2 weeks （young） and 8 weeks （adult） of age. The percentage and absolute number of NK cells in the liver and spleen were analyzed. The cytotoxicity of NK cells in the liver and spleen against various targets were detected by a 4 h ^51Cr-release method. FACScan was used to analyze the expression of CD69, Mac-l1 Ly49C/I and CD94 on the NK cells. Perforin mRNA levels were analyzed by the reverse transcription polymerase chain reaction （RT-PCR）. RESUTLS The percentages of NK cells in the liver of young and adult mice were similar （11.9%＋1.7% vs. 9.9%＋1.6%, P〉0.05）, but the absolute number per liver weight was higher in the young animals （11.6＋2.5×10^5/g vs. 3.4＋0.8×10^5/g, P〈0.05）. The level of NK cytotoxicity was extremely high in the liver of young compared to adult mice （71.0%＋5.5% vs. 23.8%＋4.4%, P〈0.05）, but this difference was not observed in the spleen. Phenotypes of the liver NK cells from young and adult mice were completely different from each other. The liver NK cells from young mice were CD69^high Mac-1^low Ly49C/ I^low, whereas NK cells from older mice displayed inverse antigen levels （CD- 69^low Mac-1^high Ly49C/I^high）. The expression levels of other NK cell-related markers were similar in both groups.The perforin mRNA level in the liver lymphocytes from young mice was consistently greater compared to adult mice. CONCLUSION From 2 to 8 weeks C57BU6 mice liver NK cells undergo age-associated changes. At 2 weeks of age the liver NK cells showed a high level of NK cytotoxicity and a unique phenotype which was not apparent at 8 weeks of age.

Analysis of Prognostic Factors of Esophageal and Gastric Cardiac Carcinoma Patients after Radical Surgery Using Cox Proportional Hazard Model-A Random Sampling Study from the Fourth Hospital of Hebei Medical University during the Period of 1996-2004

OBJECTIVE To retrospectively analyze clinical data of patientsfrom our hospital who underwent radical surgery for esophagealcarcinoma and for adenocarcinoma of the gastric cardia,as well asto investigate prognostic factors affecting the long-term survival ofthe patients.METHODS Data from the patients eligible for our study,admitted to the 4th Hospital of Hebei Medical University fromJanuary 1996 to December 2004,were randomized,and 12distinctive clinicopathologic factors influencing the survival rateof those who underwent radical surgery for esophageal carcinomaor carcinoma of the gastric cardia were collected.Univariate andmultivariate analysis of these individual variables were performedusing the Cox proportional hazard model.RESULTS It was shown by univariate analysis that age,tumorsize,pathologic type,lymph node status,TNM staging,depthof infiltration and encroachment into local organs,etc.,were thefactors that markedly influenced the prognosis of patients(P<0.01).Multivariate analysis showed that pathologic type,numberof the lymph node metastases,involvement of local organs,andTNM staging were independent prognostic factors(P<0.05).CONCLUSION The independent factors influencing theprognosis of patients with esophageal cancer and carcinoma ofthe gastric cardia include pathologic type,number of lymph nodemetastases,involvement of local organs and TNM staging.Themain prognostic factors affecting the patient's survival are patientage,tumor size and depth of infiltration.In addition,patients withinvolvement of the local organs have a worse prognosis,and theyshould be closely followed up.

Expression of Preprotachykinin-I (PPT-I), Neurokinin-1 (NK-1) and Neurokinin-2 (NK-2) in Breast Cancer Cells Improves Tumor Cell Survival in Bone Marrow in the Early Stage of Metastasis

OBJECTIVE To study the potential relationship between the expression of PPT-I, NK-1, NK2 and the development of breast cancer cells in bone marrow stroma and to provide evidence of potential molecular mechanisms of breast cancer patients. of bone metastasis in early stage METHODS The cocultures of breast cancer cell line T-47D and marrow-derived mesenchymal stem cells （MSC） were established with equal numbers. T-47D cells were separated from the coculture system at 48 h and 96 h after coculture by MACS magnetic cell sorting （MicroBeads）. The expression of PPT-I, NK-1, NK-2 in T-47D was then examined before and after coculture by real-time PCR and by Western blot. Alterations in cellular ultrastructure of T-47D cells were detected before and after coculture under electron microscope. Finally, changes in cell cycle distribution were examined by flow cytometry, and growth curves from before and after coculture were drawn and analyzed. RESULTS Following coculture of T-47D and MSC, the expression of PPT-I mRNA and protein was significantly upregulated, while the expression of NK-1 and NK-2 mRNA and protein was greatly downregulated. The analysis of cell cycle distribution by flow cytometry showed that the proportion of T-47D during S phase was increased, and the duration of the G2/M phase was sharply decreased. Under electron microscope, we observed that the synthesis of hereditary material was increased, but the hepatin granules were shown prominent stacking in T-47D cells. These results suggest that although the synthesis of DNA was increased, the proliferation of cells was inhibited after coculture. The cell growth curve confirmed the findings from the observation under the electron microscope and flow cytometry. CONCLUSION Tumor cells could survive through the upregulation in expression of preprotachykinin-I gene during early bone metastasis in breast cancer. The phenomenon of growth suppression in breast cancer cells after coculture in the current study could be induced by downregulation in expression of NK-1 and NK-2.

Investigation on complementarity between total mesorectal excision and radical resection in relation to postoperative local recurrence in patients receiving anus-reserve operation on rectal cancer

Objective:To investigate the difference and complementarity between total mesorectal excision(TME)and radical resection in relation to postoperative local recurrence in patients receiving anus-reserve operation on rectal cancer.Methods:Clinical data of 81 cases during a period from 1975 to 2001 were retrospectively analyzed.Results:In the 81 cases with local recurrence,49 of them laid to anastomosis and mesorectum,17 lymph nodes and 15 multi-site relapse.The choice of operative procedure included abdominoperineal resection in 58 cases,Hartmann’s operation in 4 cases,simple double-pelvic stoma in 12 cases,exploration in 7 cases,and total pelvic or rear-pelvic resection in combination with other organs in 6 cases. The rate of resection was 84.0%(68/81).32 cases reached clinical radical degree,and the rate of radical resection was 39.5% (32/81).The 5-year survival rate was 34.4%(11/32).Conclusion:Based on actual condition of the patients,attention to radi- cal resection and total mesorectal excision are necessary,and reasonable adoption of the operative procedure could reduce the local recurrence of rectal cancer.

Antitumor Activities of Apple Extracts

OBJECTIVE To examine the antitumor activities of fresh apple extracts. METHODS Fuji apple extracts were tested for their anti-LS^174T-pro-liferative activities, for their effect on expression of PCNA, and ability to induce apoptosis in a LS-174T cell line. RESULTS Apple extracts inhibited LS-174T cellular proliferation in a concentration and time dependent manner. The apple extracts equivalent to a concentration of 50 mg/ml inhibited the proliferation of the LS-174T cells by 34.5±1.2% after 48 h and 47.5±1.8% after 72 h respectively. Apple extracts inhibited PCNA expression and induced apoptosis of the LS-174T cells at concentrations above 12.5mg/ml. CONCLUSION Apple extracts can inhibit PCNA expression and induction of apoptosis in LS-174T cells which may contribute to their inhibitio-ry effect on cellular proliferation.

Analysis of Clinicopathologic Features of Esophageal Carcinoma Patients Undergoing Surgery-a Report of 4,329 Cases

OBJECTIVE To investigate the clinicopathological features of esophageal carcinoma （EC） patients, and to analyze epidemiologic characteristics and the current situation of esophageal cancioma in the southern area of Hebei Province. METHODS A total of 4329 patients with esophageal cancinoma, undergoing surgery in the Fourth Hospital of Hebei Medical University during a period from January 1996 to December 2005, were selected. Collection and statistical analysis of the pathologic data were performed using a SAS 6.0 software package. RESULTS Over the past ten years, there has been a tendency for an increase in the mean age of EC onset （P 〈 0.05）, a downtrend in the percentage of squamous cancer （SqCa） （P 〈 0.05） and an uptrend in the frequency of small cell carcinoma （P 〈 0.05）. In clinical stages, there was a drop in the percentage of Stage-Ⅱ squamous EC patients （P 〈 0.05）, and an increase in that of Stage-Ⅳ patients （P 〈 0.05）. There were statistical differences in sex, age, pathologic types, depth of infiltration, ratio of stages and lymph node metastasis, etc. among the superior, middle and inferior segments of the EC diseased region （P 〈 0.05）. CONCLUSION It was relatively late for the EC patients from this area to see a doctor, resulting in a drop in the ratio of SqCa and an ascensus in that of small cell cancer. However, due to a low incidence of adenocarcinoma, no obvious ascending tendency was found in the frequency of this carcinoma over the past ten years.