Comprehensive Analysis of CLCA1 Expression, Immune Infiltration and Immune Checkpoints in Colon Adenocarcinoma / Análisis Completo de la Expresión de CLCA1, Infiltración Inmune y Puntos de Control Inmunológico en Colonadenocarcinoma

SUMMARY: Colon adenocarcinoma (COAD) is a prevalent disease worldwide, known for its high mortality and morbidity rates. Despite this, the extent of investigation concerning the correlation between COAD's CLCA1 expression and immune cell infiltration remains insufficient. This study seeks to examine the expression and prognosis of CLCA1 in COAD, along with its relationship to the tumor immune microenvironment. These findings will offer valuable insights for clinical practitioners and contribute to the existing knowledge in the field. In order to evaluate the prognostic significance of CLCA1 in individuals diagnosed with colorectal cancers, we conducted a comprehensive analysis using univariate and multivariate Cox regression models along with receiver operating characteristic curve (ROC) analysis. This study was performed on the patient data of COAD obtained from The Cancer Genome Atlas (TCGA) database. Nomograms were developed to anticipate CLCA1 prognostic influence. Furthermore, the CLCA1 association with tumor immune infiltration, immune checkpoints, immune checkpoint blockade (ICB) response, interaction network, and functional analysis of CLCA1-related genes was analyzed. We found that Colon adenocarcinoma tissues significantly had decreased CLCA1 expression compared to healthy tissues. Furthermore, the study revealed that the group with high expression of CLCA1 demonstrated a significantly higher overall survival rate (OS) as compared to the group with low expression. Multivariate and Univariate Cox regression analysis revealed the potential of CLCA1 as a standalone risk factor for COAD. These results were confirmed using nomograms and ROC curves. In addition, protein-protein interaction (PPI) network analysis and functional gene enrichment showed that CLCA1 may be associated with functional activities such as pancreatic secretion, estrogen signaling and cAMP signaling, as well as with specific immune cell infiltration. Therefor, as a new independent predictor and potential biomarker of COAD, CLCA1 plays a crucial role in the advancement of colon cancer.

El adenocarcinoma de colon (COAD) es una enfermedad prevalente a nivel mundial, conocida por sus altas tasas de mortalidad y morbilidad. Sin embargo, el alcance de la investigación sobre la correlación entre la expresión de CLCA1 de COAD y la infiltración de células inmunes sigue siendo insuficiente. Este estudio busca examinar la expresión y el pronóstico de CLCA1 en COAD, junto con su relación con el microambiente inmunológico del tumor. Estos hallazgos ofrecerán conocimientos valiosos para los profesionales clínicos y contribuirán al conocimiento existente en el campo. Para evaluar la importancia de pronóstico de CLCA1 en personas diagnosticadas con cáncer colorrectal, realizamos un análisis exhaustivo utilizando modelos de regresión de Cox univariados y multivariados junto con un análisis de la curva característica operativa del receptor (ROC). Este estudio se realizó con los datos de pacientes de COAD obtenidos de la base de datos The Cancer Genome Atlas (TCGA). Se desarrollaron nomogramas para anticipar la influencia pronóstica de CLCA1. Además, se analizó la asociación de CLCA1 con la infiltración inmunitaria tumoral, los puntos de control inmunitarios, la respuesta de bloqueo de los puntos de control inmunitarios (ICB), la red de interacción y el análisis funcional de genes relacionados con CLCA1. Descubrimos que los tejidos de adenocarcinoma de colon tenían una expresión significativamente menor de CLCA1 en comparación con los tejidos sanos. Además, el estudio reveló que el grupo con alta expresión de CLCA1 demostró una tasa de supervivencia general (SG) significativamente mayor en comparación con el grupo con baja expresión. El análisis de regresión de Cox multivariado y univariado reveló el potencial de CLCA1 como factor de riesgo independiente de COAD. Estos resultados se confirmaron mediante nomogramas y curvas ROC. Además, el análisis de la red de interacción proteína- proteína (PPI) y el enriquecimiento de genes funcionales mostraron que CLCA1 puede estar asociado con actividades funcionales como la secreción pancreática, la señalización de estrógenos y la señalización de AMPc, así como con la infiltración de células inmunes específicas. Por lo tanto, como nuevo predictor independiente y biomarcador potencial de COAD, CLCA1 desempeña un papel crucial en el avance del cáncer de colon.

Construction of prognostic risk model for patients with colon adenocarcinoma based on immunogenomics analysis / 解放军医学杂志

Objective To analyze the immune genome environment of colon adenocarcinoma (COAD), find the immune genes related to the prognosis of COAD patients, and construct a prognostic risk score model to provide a basis for prognosis evaluation and individual diagnosis and treatment of COAD patients. Methods The RNA-seq data and clinical data of COAD patients were downloaded from TCGA (The Cancer Genome Atlas) database. Samples were divided into Tumor group and Normal group according to the type of tissues, and the differentially expressed immune genes were screened by R language. The immune genes related to the prognosis of COAD patients were screened by Cox regression analysis, and the prognostic risk score model was constructed. The COAD patients were divided into high-risk group and low-risk group according to their median risk score. The predictive efficiency of the immune gene prognosis model was evaluated by Kaplan-Meier analysis and Receiver Operating Characteristic (ROC) curve, and the correlation between immune gene prognostic risk model and the immune cell infiltration was analyzed. Results A total of 220 differentially expressed immune genes existed in Tumor group and Normal group. By Cox univariate and multivariate regression analysis, seven prognosis-related immune genes were screened, i.e. CXCL5 (C-X-C motif Chemokine Ligand 5), IGHV5-51 (Immunoglobulin Heavy Variable 5-51), IGKV1-33 (Immunoglobulin Kappa Variable 1-33), CHGA (Chromogranin A), UCN (Urocortin), VIP (Vasoactive Intestinal Peptide) and NR3C2 (Nuclear Receptor subfamily 3 group C member 2). The overall survival rate of COAD patients was higher in low-risk group than in high-risk group (P<0.001). The overall 5-year survival rate in high-risk group and low-risk group were 49.4% and 75.8%, respectively. ROC curve showed that AUC was 0.741, suggesting that the immune gene prognosis model had a good predictive efficiency, and was associated with immune cell infiltration of B cells, CD4+ T cells, CD8+ T cells, dendritic cells, macrophages and natural killer cells. Conclusions An immune gene risk score model has been constructed, The importance of such a prognostic model is systematically evaluated and verified in individualized treatment of patients with colon adenocarcinoma, so as to provide a direction for finding new immunotherapy targets.

G protein-coupled receptor LGR6 is an independent risk factor for colon adenocarcinoma / 医学前沿

LGR6 is a member of the G protein-coupled receptor family that plays a tumor-suppressive role in colon cancer. However, the relationship between LGR6 expression in patients and clinicopathological factors remains unclear. This study aimed to clarify whether the expression level of LGR6 is correlated with colon adenocarcinoma progression. Immunohistochemistry was used to detect LGR6 expression in colon adenoma tissues (n = 21), colon adenocarcinoma tissues (n = 156), and adjacent normal tissues (n = 124). The expression levels of LGR6 in colon adenoma and adenocarcinoma were significantly higher than those in normal colon epithelial tissues (P < 0.001). Low LGR6 expression predicted a short overall survival in patients with colon adenocarcinoma (log-rank test, P = 0.016). Univariate and multivariate survival analyses showed that, in addition to N and M classification, LGR6 expression served as an independent prognostic factor. Thus, low expression of LGR6 can be used as an independent prognostic parameter in patients with colon adenocarcinoma.

Rastreamento, diagnóstico e estadiamento do adenocarcinoma de cólon / Screening, diagnosis and staging of the colon adenocarcinoma

Introdução: O Câncer de Cólon (CC) é uma doença extremamente prevalente e letal quando não diagnosticada em tempo hábil para tratamento, o que justifica a necessidade de programas de rastreio na população. Métodos: Foram selecionados dezessete artigos das bases de dados Pubmed, Medline, Lilacs e Embase para compor uma revisão de literatura. Resultados: O objetivo do rastreamento do CC é detectar lesões precursoras (pólipos adenomatosos) ou câncer em estágio inicial, enquanto ainda são passíveis de tratamento. Pode-se utilizar diversos métodos para realizar este rastreamento, sendo a colonoscopia o exame padrão-ouro devido a capacidade de estabelecer diagnóstico histopatológico da doença e fornecer oportunidade de tratamento no mesmo procedimento em casos de lesão precursoras. O estadiamento é clínico e envolve exame físico, radiografia torácica e tomografia computadorizada (TC) de abdome e pelve. Conclusão: O CC é uma das neoplasias mais frequentes em adultos no mundo inteiro e é possivelmente curável se diagnosticado precocemente. O método padrão-ouro para rastreamento e diagnóstico é a colonoscopia, uma vez que ela permite biópsia para análise histopatológica na presença de lesão. O estadiamento deve ser feito para avaliar a extensão local e sistêmica da doença por meio do exame físico e exames complementares.

Introduction: The colorectal cancer (CC) is an extremely common disease that is lethal if not diagnosed in early stages, which justifies the need for screening in the general population. Methods: Seventeen publications from Pubmed, Medline, Embase and Lilacs databases were selected to compose a literature review. Results: The goal of screening for CC is to detect precursor lesions (polyps) or early stage cancer. There is a variety of methods available in order to perform this screening, being colonoscopy the gold standard due to the possibility of establishing both histopathological diagnosis of the disease and offering treatment in case of precursor lesions. The staging is clinical and includes physical examination, chest imaging and abdominal and pelvic CT scans. Conclusion: The CC is one of the most common cancers in adults and possibly treatable if early diagnosed. The gold standard method for screening is colonoscopy, since it allows biopsy for histopathological analysis. The staging needs to be done to assess the local and systemic extent of the disease through physical examination and complementary tests.

Is the Location of the Tumor Another Prognostic Factor for Patients With Colon Cancer?

PURPOSE: In this study, we investigated both the characteristics of right colon cancer (RTCC) in comparison with those of left colon cancer (LTCC) and the impact of the location of the colon cancer on the prognosis. METHODS: We retrospectively analyzed the cases of 974 patients with nonmetastatic colon cancer who had undergone surgery with a curative intent from January 2001 to December 2011. RTCC was defined as a tumor located proximal to the splenic flexure. The characteristics of RTCC cancer were investigated by using descriptive analyses, and their impacts on the prognosis were assessed by using a Cox multivariate regression. RESULTS: Compared to LTCC, RTCC showed a female-dominant feature, and an undifferentiated pathology was more frequently observed. The number of lymph nodes retrieved from patients with RTCC was significantly higher than that retrieved from patients with LTCC. During 75 months of follow-up, peritoneal recurrence was more common in patients with RTCC than it was in patients with LTCC, and among the patients with stage III colon cancer, the disease-free and the overall survival rates were significantly worse in patients with RTCC. After adjustments with the other prognostic factors associated with colon cancer had been made, a tumor located at the right colon was found to be independently associated with poor prognosis. CONCLUSION: RTCC showed unique clinicopathologic features and was associated with a poorer prognosis.

Tumor sincrónico: cáncer de colon y liposarcoma / Synchronous tumor: colon cancer and liposarcoma

Se presenta el caso clínico de un paciente varón de 58 años, natural de Junín, que ingresó por Emergencia, referido de un hospital provincial por un cuadro de obstrucción intestinal, con un tiempo de enfermedad de 7 días, caracterizado por dolor abdominal tipo cólico difuso, náuseas y vómitos y distensión abdominal. Por dos meses presentaba dolor abdominal intermitente intenso, 3 a 4 episodios por mes, que cedía en forma espontánea, pero progresivamente aumentó su frecuencia y se asoció con ingesta de alimentos, disminuyó el apetito y perdió peso aproximadamente 6 a 7 kilos; no presentaba antecedentes patológicos ni quirúrgicos de importancia. Durante su hospitalización en nuestro nosocomio se obtuvo una tomografía (TAC) abdominal que mostró la presencia de dos tumoraciones, una en colon ascendente y otra en región retroperitoneal, que fueron extraídas en cirugía electiva. El resultado de la anatomía patológica fue adenocarcinoma bien diferenciado de colon y liposarcoma dediferenciado del tumor retroperitoneal; ambos presentaron bordes quirúrgicos libres de neoplasia. La coincidencia de ambas patologías no ha sido descrita en la literatura médica.

This is a case report of a 58 year-old male born in Junin. Patient was referred to the emergency room from a rural hospital and was admitted with the diagnosis of intestinal obstruction. He had colicky type diffuse abdominal pain, abdominal distention, nausea and vomiting for 7 days. He had presented abdominal pain off and on for the past 2 months. Pain was severe at least 3 to 4 times a week, improved spontaneously but returned with increased frequency, exacerbated by food ingestion, and associated with decreased appetite and 6 to 7 kg weight loss. Patient did not have any significant past medical or surgical history. During hospitalization abdominal CT showed two tumors, one in the ascending colon and the other in the retroperitoneal region. These were surgically removed. Pathology result reported well differentiated colon adenocarcinoma and dedifferentiated liposarcoma, both with surgical borders clean of neoplasia. Simultaneous finding of these tumors has not been described in the medical literature.

Sigmoidoanal intussusception with exteriorization of sigmoid adenocarcinoma

The intestinal intussusception is a rare disease in adults, and is mostly caused by malignant neoplasm. Symptoms are usually nonspecific and chronic, and in most cases suggesting intestinal obstruction. Treatment consists of removing the malignant tumor. This article reports the case of a patient with hematochezia and apparent mass in the anus who underwent anterior rectosigmoidectomy and had the diagnosis of adenocarcinoma of the sigmoid confirmed. (AU)

A intussuscepção intestinal é uma doença rara em adultos, sendo na maior parte dos casos causada por neoplasia maligna. Os sintomas são geralmente inespecíficos e crônicos, na maioria das vezes sugerindo obstrução intestinal. O tratamento consiste na remoção oncológica do tumor. Este artigo relata o caso de uma paciente com quadro de hematoquezia e exteriorização de massa através do ânus que foi submetido à retossigmoidectomia anterior alta em bloco e confirmado o diagnóstico de adenocarcinoma de sigmoide. (AU)

Synchronous Incidental Occurrence of Gastric Gastrointestinal Stromal Tumor and Colon Adenocarcinoma / 대한소화기내시경학회지

A few cases of gastrointestinal stromal tumor (GIST) synchronous cancers with other malignancies have been reported, such as gastric cancer, pancreatic cancer, renal cell carcinoma, colon cancer, and carcinoid tumor. However, little is known about their coincidence with other tumors with a different histogenesis. A 62-year-old man visited our hospital with generalized weakness and intermittent hematochezia. A colonoscopic examination showed an ulcerating mass in the ascending colon, and a biopsy specimen revealed adenocarcinoma. A lobulated submucosal mass in the gastric fundus was found incidentally during the preoperative staging procedures, including computed tomography (CT) and positron emission tomography-CT. The colon cancer and gastric tumor were removed simultaneously (laparoscopy assisted right hemicolectomy and total gastrectomy). Immunohistochemical studies on the gastric tumor surgical specimen showed that it was CD117 (+) and CD34 (+), and the final diagnosis was a GIST of the stomach. Herein, we report a case of the synchronous occurrence of colon adenocarcinoma and gastric GIST.

Immunohistochemical Studies of Fascin, MMP-9 Overexpression, and Proliferating Index as Prognostic Factors in Cases of a Colon Adenocarcinoma

PURPOSE: The malignant conversion of epithelial cells involves alterations in the expression and the function of cell-matrix and cell-cell adhesive systems that enable a switch to a migratory phenotype in tumor invasion and metastasis. Here, the author studies the prevalence and the potential clinical significance of fascin and Matrix metalloproteinase-9 (MMP-9) expression in relation to the progression of colon adenocarcinoma and of tumor cell proliferation as measured by using the topoisomerase II-alpha (Topo II-alpha) index. METHODS: Relatively well-preserved paraffin-embedded tissues of 120 cases of colon adenocarcinomas were immunohistochemically stained for fascin, MMP-9, and Topo II-alpha expression. A reaction was determined as being positive when more than 10% of the cells were positive for fascin, and/or MMP-9. The Topo II-alpha index is defined as the positive number of tumor cells divided by the total number of tumor cells counted times 100. At least 1,000 cells were counted for this analysis. A chi-square test, by using Epi info 2000, for Fascin and/or MMP-9 and a two-sided test for the Topo II-alpha index were employed with a significance of P65 yr, P=0.028), tumor grading (P=0.009), and lymph node metastases (P=0.005). However, MMP-9 immunoreactivity was not statistically associated with age, gender, tumor stage, or lymph node metastases. Fascin expression was statistically associated with MMP-9 expression, especially for left colon adenocarcinomas (P=0.0032). Although the topo II-alpha proliferating index was associated with lymph node metastasis (P<0.01), this result was not statistically associated with Fascin or MMP-9 expression. CONCLUSION: Fascin expression may be closely linked with tumor grading and lymph node metastasis of more aggressive colon adenocarcinomas and partly associated with MMP-9 expression in tumor invasion. However, further studies of fascin expression as an independent prognostic factor are required for the determination of significant relationships with other clinicopathologic indices.

A Case of Cutaneous Metastases to the Face and Scalp from Colon Adenocarcinoma / 대한피부과학회지

Cutaneous metastases occur uncommonly with a reported incidence of 0.7% to 10% in all patients with internal organ malignancy. Cutaneous metastases in the face occur in less than 0.5% of patients with metastatic skin cancer. Cutaneous metastases occur in less than 4% of colon adenocarcinoma patients and the most frequent site is the abdominal skin, especially the scar of previous surgical incision. But metastases to the skin of the head and neck region from colon adenocarcinoma are rare and especially, facial skin metastases are very rare. We report a case of cutaneous metastasis to the face and scalp from colon adenocarcinoma.

Estudo da expressão citofotométrica do marcador tumoral Caspase-3 no adenocarcinoma de cólon / Citophotometric expression study of tumoral marker Caspase-3 on colon adenocarcinoma

RACIONAL: O adenocarcinoma de cólon é a segunda causa mais comum de morte por câncer em homens e mulheres, sendo responsável por mais de cinco milhões de mortes por ano. No momento do diagnóstico apenas 70% dos tumores são ressecáveis, 75% são curáveis e 25% poderão ter recorrência da doença. A apoptose é uma das responsáveis pelo equilíbrio homeostático entre as células. Durante o desenvolvimento do processo de degeneração maligna celular o desequilíbrio na apoptose é considerado um dos principais marcos neoplásicos. A caspase-3 é uma das mais importantes moléculas na apoptose, sendo sua efetora principal. Sua expressão e prognóstico têm sido relatados em vários estudos e revisões com seu papel valorizado desde o surgimento do pólipo até a sua transformação maligna, com a taxa de apoptose diminuindo progressivamente. OBJETIVOS: Avaliar a expressão citofotométrica computadorizada do marcador Caspase-3 no adenocarcinoma de cólon; avaliá-lo nas fases evolutivas na classificação modificada de Dukes e comparar sua expressão nos tumores do lado direito e esquerdo do cólon. MÉTODOS: Utilizaram-se 19 casos de câncer recuperados de blocos de parafina confirmados por hematoxilina-eosina e submetidos à técnica imunoistoquímica da estreptavidina-biotina com anticorpo policlonal anti-caspase-3. Após este processo as lâminas marcadas foram submetidas à leitura pelo sistema SAMBA com o software IMUNNO 4.00. Foram analisados três índices: marcagem (Label index), heterogeneidade e densidade óptica. Utilizaram-se a marcagem individual, avaliação da expressão do marcador e grupos definidos de tumores com classificação Dukes e pelo lado do tumor. RESULTADOS: A média do índice de marcagem da caspase-3 foi de 85,24 e da densidade óptica de 39,55. Na classificação Dukes de 12 tipos B tiveram índice de marcagem de 86,20 e a densidade óptica de 37,72 e para os 7 tipos C a área de marcagem foi de 85,66 e a densidade óptica foi de 42,71 não sendo possível identificar diferença em relação a classificação de Dukes. Quanto ao lado do tumor os 11 tumores à esquerda tiveram índice de marcagem de 86,65 e densidade óptica de 43,29 e os 8 à direita tíndice de marcagem de 83,29 e densidade óptica de 39,44 não sendo possível observar diferença estatística significante. CONCLUSÕES: A caspase-3 possui alta expressão individual revelando-se marcador de boa utilidade no estudo do adenocarcinoma de cólon e da fase pró-apoptóica da sua tumorigênese pelo seu alto grau de índice de marcagem e densidade óptica. Em relação à classificação Dukes não houve diferença entre os tipos B e C, como também em relação ao lado direito e esquerdo do cólon.

BACKGROUND: Colon adenocarcinoma is the second cause of cancer death in men and women, responsible for more than five million deaths each year. At the diagnosis, only 70% of tumors are resectable, 75% curative and 25% might have disease recurrence. Apoptosis is responsible for homeostatic equilibrium among cells. During the development of the cellular malignant degeneration, an abnormal apoptosis plays an important role in the neoplasm transformation. Caspase-3 is one of the most important molecules in apoptosis, considered as the principal executioner. Caspase-3 expression and its prognosis have been mentioned in a variety of study and revision as its role being emphasized since the onset of polyp formation until the malignant transformation, as apoptosis declines progressively. AIM: To evaluate the computerized image cytophotometric expression of Caspase-3 in colon adenocarcinoma; to evaluate in the respective phases of Dukes-modified Classification and to compare its expression on right-sided and left-sided colon tumors. METHODS: Nineteen recovery of sampled-tissue in paraffin blocks confirmed by hematoxylin-eosin and submitted to immunohistochemistry estreptavidin-biotin method using antibody polyclonal anti-caspase-3. Following this process, marked slides were submitted SAMBA reading using software IMUNNO 4.00. Three indexes were analyzed: label index, heterogeneity and optical density. Individual labeling, marker expression evaluation and predefined tumors groups under Dukes Classification and side of tumor occurrence were considered. RESULTS: The average labeling index for caspase-3 was 85,24, where as for optical density was 39,55 pixels. According to Dukes Classification, 12 type B presented labeling index of 86,20 and optical density of 37,72 and for 7 samples type C labeling index corresponded to 85,66 and optical density equals to 42,71, unable to identify the difference regarding to Dukes Classification. Regarding to the side of tumor occurrence, 11 left-sided tumors had labeling index of 86,65 and optical density of 43,29 and the 8 right-sided tumors had labeling index of 83,29 and optical density of 39,44, unable to observe significant statistical difference. CONCLUSION: Caspase-3 presents a distinct value expression, revealing as useful marker in colon adenocarcinoma study and its proapoptotic phase at the tumorigenesis due to its high labeling index and optical density. Regarding to Dukes Classification there was no difference between types B and C, as well as right and left sided-related colon tumor occurrence.

Two Cases of Colon Adenocarcinoma Transformed from Colonic Adenoma / 대한소화기내시경학회지

The prevalence of colorectal cancer is increasing now in Korea and the principal strategy of its management is early detection and surgical resection. Current discussion of the topic of colorectal polyps and cancer is largely based on the concept of the adenoma-carcinoma sequence, which is thought to be the most probable pathogenesis for colorectal cancer. But the case reports about colonic adenocarcinoma arising from adenoma in clinical practice between short interval are rare in Korea. We experienced two cases of colon cancer, which were transformed from small adenoma can not be removed because of patients' refusal, 3 years ago. So we present these cases with a review of relevant literatures.

Roles of T Cells and Related Immunological Molecules in the Enhancement of Cancer CD Suicide Gene Therapy by Lymphotactin / 中国肿瘤生物治疗杂志

We previously showed that adenvirus-mediated lymphotactin (Ltn) gene transfer in vivo could improve the an-titumor efficacy of cytosine deaminase (CD) gene therapy significantly. In the precent study, we investigated the im-munological mechanisms involved in the enhanced antitumor efficacy. Upregulation of CD80 and CD54 on murine CT26 colon carcinoma cells was observed after combined transfection with adenovirus encoding CD (AdCD) and adenovirus encoding murine Ltn ( AdLtn) followed by administration of 5-PC in vitro. IL-2 and IFN-? level secreted by splenocytes increased significantly after the combination therapy. In vivo depletion analysis showed that both CD4~+ and CD8~+ T cells participated in the antitumor effect of the host with CD8~+ T cells being the main T cell subset responsible for the enhanced antitumor immune response. These data suggested that increased irnmunogenicity and efficient induction of antitumor immunity of the host might contribute to the enhanced antitumor effects of the combined Ltn and CD suicide therapy.

Antitumor Effects of G-CSF Gene Therapy on Colon Adenocarcinoma-Bearing Mice Receiving High-Dose Chemotherapy / 中国肿瘤生物治疗杂志

Granulocyte colony - stimulating factor (G - CSF) is a hematopoietic growth factor that is responsible for the differentiation and proliferation of hematopoietic progenitor cells to mature granulocyte, and can increase the number of peripheral neutrophils. It has been demonstrated that it could inhibit the metastasis of the murine tumors in spontaneous and experimental metastasis models by in vivo administration of recombinant human G - CSF. In order to examine the antitumor effect of G - CSF gene therapy on mice receiving high - dose chemotherapy, C - 26 colon adenocarcinoma - bearing mice which were prepared by S. c. injection of 1?105C-26 cells were i. p. injected with rhG-CSF (2?g/day?14day) or implanted with 1?107 collagen encapsulated NIH3T3-G-CSF cells which secrete high level of G - CSF after gene transfection. In our experiment, rhG-CSF could inhibit the tumor growth and extend the survival time of early stage C-26 bearing mice. However, G - CSF gene therapy could inhibit the tumor growth and prolong the survival both in early or middle stage C-26 mice. The results showed that both rhG - CSF and G - CSF gene therapy have exact antitumor effect and G - CSF gene therapy show more effective than rhG - CSF in vivo. Then we investigated the therapeutic effects of G - CSF gene therapy on C-26-bearing mice receiving high - dose chemotherapy (5-Fu 150mg/mice i. p.) . More effective results could be observed in C-26 - bearing mice receiving high dose chemotherapy after G - CSF gene therapy. The results also suggested that G-CSF gene therapy can inhibit the tumor growth more effectively both in C-26-bearing mice or C-26-bearing mice receiving high - dose chemotherapy.

Effects of G-CSF Gene Therapy bn the Number and Functions of Peripheral Neutrophils of Colon Adenocarcinoma-Bearing Mice Receiving High-Dose Chemotherapy / 中国肿瘤生物治疗杂志

Neutrophils play important role in anti - tumor response of tumor-bearing host as a kind of important effector cells. In order to identify the anti-tumor mechanisms of G-CSF gene therapy, we investigated the number and functions of the peripheral neutrophils in the C-26 colon adenocarcinoma-bearing mice receiving G - CSF gene therapy and high - dose chemotherapy. After G - CSF gene therapy, the number of neutrophils in peripheral blood was increased markedly in C-26 mice receiving high - dose 5 - Fu as compared with control groups including in vivo administration of rhG - CSF. The more potent cytotoxicity to C - 26 cells could be detected. The phagocytic activity, the secretion of IL-1, TNF, NO of the neutrophils were significantly enhanced. These data showed that G - CSF gene therapy can increase the number of neutrophils, activate the functions more effectively than in vivo administration of rhG - CSF.

Combined Use of CD Suicide Gene and Lymphotactin Gene Therapy Elicit Potent Antitumor Effects / 中国肿瘤生物治疗杂志

Adenovirus harboring E. coli. cytosine deaminase gene (AdCD) and adenovirus encoding with lymphotactin gene (AdLtn) were used for gene therapy in vivo. BALB/c mice were inoculated subcutaneously with CT26 colon adeno-carcinoma cells and 3 days later received combined injection of AdCD and/or AdLtn followed by continuous injection with 5-fluorocytosine(5-FC) 300mg/kg. The results demonstrated that mice received combined therapy developed tumors most slowly and survived longest when compared with mice treated with AdCD/5-FC, AdLtn, AdlacZ/5-FC or PBS. To further explain the immunological mechanism of the antitumor effects by the combined therapy, we found that combined treatment with suicide gene and Ltn gene therapy achieved maximal cytotoxic effects of nature killer cells and specific cy-totoxic T lymphocytes. FACS analysis of the tumor mass demonstrated that AdCD/5-FC in combination with AdLtn therapy increased the expression of H-2K~d and B7-1 expression on tumor cells. The CD4~+ and CD8~+ cells infiltrated in the tumor mass after combined therapy were significantly increased when measured by FACS analysis. Our results demonstrated that combined transfer of suicide gene and lymphotactin gene induce nonspecific and specific antitumor immunity of the host and elicit more potent antitumor effect.

Interpretation of DNA Histogram in Flow Cytometry: A Comparative Study of DNA Ploidy in Fresh and Paraffin-embedded Tissues of Colorectal Adenocarcinomas

As flow cytometric analysis using paraffin-embedded tissue was developed by Hedley et al in 1983, retrospective study with large amount of archival material was possible. Many literatures reported that the result of paraffin embedded tissue was compatible with that of fresh tissue. We compared the DNA histograms of 26 cases of colorectal adenocarcinoma in which the analysis was done in both fresh and paraffin-embedded tissues. Aneuploidy in fresh and paraffin-embed-ded tissues was 73.0% and 50.0%, respectively. The concordance rate of fresh and paraffin-em-bedded tissues was 76.8% and six interpreters were agreed in 73.0% of the cases. Because flow cytometric DNA analysis using fresh tissues can detect more aneuploid population than in paraffin-embedded tissue, the former is strongly recommeded in DNA ploidy study. Also careful observation using standard criteria may improve the interpretation of DNA histogram.