Effects of splenic lymphocytes isolated from the mouse of colorectal carcinoma with liver metastases induced by oncolytic herpes simplex virus type Ⅱ on colorectal carcinoma with pulmonary metastases in mice / 肿瘤研究与临床

Objective:To study the effect of splenic lymphocytes isolated from mouse models of colorectal carcinoma with liver metastases induced by oncolytic herpes simplex virus type Ⅱ(oHSV2) on the growth of pulmonary metastases of colorectal carcinoma.Methods:A total of 18 6-week-old BALB/c female mice were selected, colorectal carcinoma cell line CT26 of mice in logarithmic phase was inoculated at the right back (2×10 5 per mouse) and spleen (1×10 5 per mouse) of mice, and tumor cells had hematogenous metastasis to liver through splenic vein. CT26 colorectal carcinoma with liver metastases model was constructed. All mice were respectively divided into oHSV2 group and phosphatic buffered saline (PBS) group, 9 mice in each group according to the random number table method. Mice in oHSV2 group were treated with subcutaneous intratumoral multi-point injection of 100 μl oHSV2 (the multiplicity of infection was 1) for 6 cycles, while mice in PBS group were treated with subcutaneous intratumoral multi-point injection of 100 μl PBS for 6 cycles in total, once injection every other day; the survival of mice was analyzed by using Kaplan-Meier method and tumor growth was observed. The mice of both groups in mouse models of colorectal carcinoma with liver metastases were killed on day 20 and their splenic lymphocytes were isolated. After investigation of the most suitable inoculation number and the optimal observation time of colorectal carcinoma with pulmonary metastases CT26 cell lines, 9 6-week-old BALB/c female mice were divided into the experimental group, the negative control group and the blank control group according to the random number table method, with 3 mice in each group. Mice in the experimental group were injected with splenic lymphocytes (4×10 7 per mouse) and CT26 cells (2×10 5 per mouse) isolated from mouse models induced by oHSV2 via the tail vein, mice in the negative control group were injected with splenic lymphocytes (4×10 7 per mouse) and CT26 cells (2×10 5 per mouse) isolated from normal mice with same weeks old via the tail vein, and mice in the blank control group were injected with only CT26 cells (2×10 5 per mouse) via the tail vein. The above 3 groups were executed on day 10 after inoculation, and tumor growth, histopathological changes of mice were also observed; the survival of mice was analyzed by using Kaplan-Meier method. Results:In models of colorectal carcinoma with liver metastases, liver metastases lesions were not detected in 7 mice and 2 mice had 1-2 liver metastases lesions with long diameter less than 2 mm of oHSV2 group; in PBS group, 9 mice all had multiple liver metastases lesions with tumor long diameter ranging from 1 to 10 mm. And mice in oHSV2 group survived much longer than that of mice in PBS group ( P < 0.001). In models of pulmonary metastases, the optimal number of CT26 cells in mouse tail vein was 2×10 5 per mouse; the best observation time point was day 10 after tail vein injection. On day 24 after inoculation, all mice in the negative control group and the blank control group died, while mice in the experimental group all survived on day 60, and the difference of the overall survival in the above 3 groups was statistically significant ( P = 0.007). HE staining results showed that the lung tissues of the experimental group did not show clear tumor cells, whereas the lung tissues of the negative control group and the blank control group showed extensive diffuse tumor cells. Conclusions:Splenic lymphocytes produced by oHSV2 induction in mouse models of colorectal carcinoma with liver metastases can effectively inhibit the development of pulmonary metastases in colorectal carcinoma CT26 cell of mice.

Role of heat-shock proteins in infection of human adenocarcinoma cell line MCF-7 by tumor-adapted rotavirus isolates / Papel de las proteínas de choque térmico en la infección de la línea celular de adenocarcinoma humano MCF-7 por aislados de rotavirus adaptados a tumores

Abstract Background: Viruses are being used as alternative and complementary tools for treating cancers. Oncolytic viruses exhibit tumor tropism, ability to enhance anti-tumor immunity and ability to be used in combination with conventional chemotherapy and radiotherapy. We have recently selected some rotavirus isolates which are adapted to efficiently infect and kill tumor cell lines. Aim: We tested five tumor cell-adapted rotavirus isolates for their ability to infect the human adenocarcinoma cell line MCF-7. Methods: Cell surface membrane-associated proteins mediating virus particle attachment were characterized using ELISA, immunoprecipitation, FACS analysis, and antibody blocking. Results: It was found that heat shock proteins (HSPs) such as Hsp90, Hsp70, Hsp60, and Hsp40 are expressed on the cell surface forming complexes with protein disulfide isomerase (PDI), integrin β3, and heat shock cognate protein 70 (Hsc70) in lipid raft microdomains. Interaction of rotavirus isolates with these cellular proteins was further confirmed by a competition assay and an inhibition assay involving the HSPs tested. Conclusion: Our findings suggest that the tumor cell-adapted rotavirus isolates studied here offer a promising tool for killing tumor cells, thus encouraging further research into this topic, including animal models.

Resumen Antecedentes: Los virus se utilizan como herramientas alternativas y complementarias para el tratamiento del cáncer. Los virus oncolíticos exhiben tropismo por tumores, capacidad para intensificar la inmunidad antitumoral y la capacidad para utilizarse en combinación con quimioterapia y radioterapia convencionales. Recientemente, hemos seleccionado algunos aislamientos de rotavirus que están adaptados para infectar y eliminar de manera eficiente líneas de células tumorales. Objetivo: Se ensayaron cinco aislamientos de rotavirus adaptados a células tumorales para determinar su capacidad para infectar la línea celular de adenocarcinoma humano MCF-7. Métodos: Las proteínas asociadas a la membrana de la superficie celular que median la unión de partículas de virus se caracterizaron mediante ELISA, inmunoprecipitación, análisis FACS y bloqueo de anticuerpos. Resultados: Se encontró que las proteínas de choque térmico (HSPs) como Hsp90, Hsp70, Hsp60 y Hsp40 se expresan en la superficie celular formando complejos con la proteína disulfuro isomerasa (PDI), la integrina β3 y la proteína análoga de choque térmico 70 (Hsc70) en microdominios lipídicos (rafts). La interacción de los aislamientos de rotavirus con estas proteínas celulares se confirmó adicionalmente mediante un ensayo de competición y un ensayo de inhibición que incluía las HSP ensayadas. Conclusión: Nuestros hallazgos sugieren que los aislamientos de rotavirus adaptados a las células tumorales estudiados aquí ofrecen una herramienta prometedora para eliminar las células tumorales, lo que estimula más investigaciones sobre este tema, incluidos los modelos animales.

Assessing the oncolytic potential of rotavirus on mouse myeloma cell line Sp2/0-Ag14 / Evaluación del potencial oncolítico del rotavirus en la línea celular Sp2/0-Ag14 de mieloma de ratón

Introduction: Cancer is the second leading cause of death in the United States, surpassed only by cardiovascular disease. However, cancer has now overtaken cardiovascular disease as the main cause of death in 12 countries in Western Europe. The burden of cancer is posing a major challenge to health care systems worldwide and demanding improvements in methods for cancer prevention, diagnosis, and treatment. Alternative and complementary strategies for orthodox surgery, radiotherapy, and chemotherapy need to be developed. Objective: To determine the oncolytic potential of tumor cell-adapted rotavirus in terms of their ability to infect and lysate murine myeloma Sp2/0-Ag14 cells. Materials and methods: We inoculated rotaviruses Wt1-5, WWM, TRUYO, ECwt-O, and WTEW in Sp2/0-Ag14 cells and we examined their infectious effects by immunocytochemistry, immunofluorescence, flow cytometry, and DNA fragmentation assays. Results: Rotavirus infection involved the participation of some heat shock proteins, of protein disulfide isomerase (PDI), and integrin ß3. We detected the accumulation of viral antigens within the virus-inoculated cells and in the culture medium in all the rotavirus isolates examined. The rotavirus-induced cell death mechanism in Sp2/0-Ag14 cells involved changes in cell membrane permeability, chromatin condensation, and DNA fragmentation, which were compatible with cytotoxicity and apoptosis. Conclusions: The ability of the rotavirus isolates Wt1-5, WWM, TRUYO, ECwt-O, and WTEW to infect and cause cell death of Sp2/0-Ag14 cells through mechanisms that are compatible with virus-induced apoptosis makes them potential candidates as oncolytic agents.

Introducción. El cáncer es la segunda causa de muerte en los Estados Unidos, solamente superado por la enfermedad cardiovascular. Sin embargo, el cáncer aventaja a la enfermedad cardiovascular como primera causa de muerte en doce países de Europa occidental. Se requieren mejores métodos de prevención, diagnóstico y tratamiento para afrontar el gran desafío que el cáncer representa mundialmente para los sistemas de salud, y se necesita desarrollar estrategias alternativas y complementarias a la cirugía, la radioterapia y la quimioterapia convencionales. Objetivo. Evaluar el potencial oncolítico de rotavirus adaptados a células tumorales por su capacidad para infectar y lisar células Sp2/0-Ag14 de mieloma de ratón. Materiales y métodos. Los aislamientos de rotavirus Wt1-5, WWM, TRUYO, ECwt-O y WTEW se inocularon en células Sp2/0-Ag14 y se examinaron sus efectos infecciosos mediante inmunocitoquímica, inmunofluorescencia, citometría de flujo y ensayos de fragmentación del ADN. Resultados. La infección con los rotavirus Wt1-5, WWM, TRUYO, ECwt-O y WTEW implicó la participación de algunas proteínas de choque térmico, la proteína disulfuro isomerasa y la integrina ß3. La acumulación de antígenos virales intracelulares y extracelulares se detectó en todos los virus utilizados. Los mecanismos de muerte inducidos por los rotavirus en células Sp2/0-Ag14 indujeron cambios en la permeabilidad de la membrana celular, la condensación de cromatina y la fragmentación de ADN, los cuales fueron compatibles con citotoxicidad y apoptosis. Conclusiones. La capacidad de los rotavirus estudiados para infectar y causar la muerte de células Sp2/0-Ag14 mediante mecanismos compatibles con la apoptosis inducida viralmente los convierte en candidatos potenciales para ser utilizados como agentes oncolíticos.

Virus oncolíticos: un arma contra el cáncer / Oncolytic viruses: a weapon against cancer

Resumen Introducción. Los virus oncolíticos son virus atenuados, mutados o que por naturaleza se dirigen y matan específicamente células tumorales, sin afectar a las células normales. La administración intratumoral del virus ofrece la oportunidad de tratar el tumor primario pero no focos metastásicos, los cuales pueden ser alcanzados mediante la administración intravenosa. Sin embargo, su eficiencia puede disminuir por la presencia de una respuesta inmunológica preexistente en los sujetos tratados. Objetivo. Exponer las técnicas utilizadas para envolver y transportar los virus con el fin de eludir el sistema inmunológico antes de que el virus llegue al tumor. Materiales y métodos. Se realizó una búsqueda narrativa de la literatura original y de revisión en las bases de datos PubMed, JSTOR y EBSCO sobre métodos o técnicas utilizadas para el tratamiento del cáncer mediante el uso de virus oncolíticos. Resultados. La formación de nanocomplejos entre los virus oncolíticos y biopolímeros -ya sea mediante la unión química o mediante la unión a través de interacciones electrostáticas o el uso de micropartículas, células transportadoras, liposomas, ultrasonido o terapias combinadas- es eficaz para evitar la respuesta inmunológica del huésped contra el virus. Conclusión. Para evitar la respuesta inmunológica del huésped contra los virus oncolíticos se han desarrollo diversos métodos que permiten la liberación controlada y especifica de los mismos. Sin embargo, debido a la diversidad de los virus, se debe tener en cuenta que la eficacia de los métodos de protección y transporte depende de las características bioquímicas tanto del biomaterial como del virus.

Abstract Introduction: Oncolytic viruses are attenuated, mutated, or naturally ocurring viruses that specifically kill tumor cells without affecting normal cells. Intratumoral administration of the virus offers the opportunity to treat the primary tumor but not metastatic foci, which can be done through intravenous administration. However, its efficacy may be reduced by the presence of a pre-existing immune response in treated subjects. Objective: To present the techniques used to wrap and transport viruses in order to bypass the immune system before the virus reaches the tumor. Materials and methods: A narrative search of original and review literature was conducted in the PubMed, JSTOR and EBSCO databases on methods or techniques used for the treatment of cancer using oncolytic viruses. Results: The formation of nanocomplexes between oncolytic viruses and biopolymers -either by chemical binding or electrostatic interactions, or cell-derived microparticles, carrier cells, liposomes, ultrasound or combination therapies- is effective in preventing the host's immune response against the virus. Conclusion: Different methods that depend on the type of oncolytic virus have been developed to increase the efficacy of the therapeutic response. Controlled and specific-release virus delivery systems have been developed to avoid the immune response against them. However, due to the diversity of viruses, it should be borne in mind that the effectiveness of protection and transport methods depends on the biochemical characteristics of both the biomaterial and the virus.

Progress of anti-tumor mechanisms of reovirus / 肿瘤研究与临床

Oncolytic viruses have made great breakthroughs in cancer treatment, especially for reovirus, which can effectively induce the death of tumor cells without harming the normal tissues. More than 80％ tumor cells are sensitive to reovirus infection. Reovirus induces the apoptosis of tumor cells and exerts anti-tumor immunity to achieve anti-tumor activity, and the curative effect of combination therapy with reovirus and chemotherapeutic drugs exceeds the effect of monotherapy. Reovirus can exert anti-tumor effects through different mechanisms, which is of great significance for the new and effective treatment of tumors in the future.

Cell Death Mechanisms in Esophageal Squamous Cell Carcinoma Induced by Vesicular Stomatitis Virus Matrix Protein

OBJECTIVES: Vesicular stomatitis virus (VSV) is under development as an oncolytic virus due to its preferential replication in cancer cells and oncolytic activity, however the viral components responsible have not yet been determined. In this study the effects of VSV wild-type (wt) and M51R-mutant matrix proteins (M51R-mMP) on apoptosis, pyroptosis, necroptosis, and autophagy pathways, in an esophagus cancer cell line (KYSE-30) were investigated. METHODS: The KYSE-30 cells were transfected with pcDNA3.1 plasmids encoding wt or M51R-mMP, and apoptosis, pyroptosis, necroptosis, and autophagy were evaluated 48 and 72 hours after transfection. RESULTS: KYSE-30 cells transfected with VSV wt and M51R-mMPs significantly reduced cell viability to < 50% at 72 hours post-transfection. M51R-MP significantly increased the concentration of caspase-8 and caspase-9 at 48 and 72 hours post-transfection, respectively ( p < 0.05). In contrast, no significant changes were detected following transfection with the VSV wt plasmid. Moreover, VSV wt and M51R-mMP transfected cells did not change the expression of caspase-3. VSV wt and M51R-mMPs did not mMP change caspase-1 expression (a marker of pyroptosis) at 48 and 72 hours post-transfection. However, M51R-mMP and VSV wt transfected cells significantly increased RIP-1 (a marker of necroptosis) expression at 72 hours post-infection ( p < 0.05). Beclin-1, a biomarker of autophagy, was also induced by transfection with VSV wt or M51R-mMPs at 48 hours post-transfection. CONCLUSION: The results in this study indicated that VSV exerts oncolytic activity in KYSE-30 tumor cells through different cell death pathways, suggesting that M51R-mMP may potentially be used to enhance oncolysis.

Immunotherapy for colorectal cancer: current status, challenges and solution / 中国肿瘤生物治疗杂志

@# Colorectal cancer (CRC) is the third most common malignancy and the fourth leading cause of cancer-related death all over the world. Traditional treatments, including chemotherapy, radiation therapy, surgery and targeted therapy, form the backbone of current treatment in various stages of CRC, but the efficacy in patients with recurrent or metastatic disease is extremely poor. Recently-developed immunotherapy is frequently used in various cancers with high malignancy, such as leukemia, melanoma and non-small-cell lung cancer, and achieves promising clinical outcomes. Immunotherapy has been also investigated in CRC, but the outcome is so disappointed in majority of patients, except the PD-1 inhibitor achieved excellent result in CRC with DNA mismatch repair system deficiency. In this review, the authors will mainly introduce the immunophenotype of different subtype of CRC and summarize current advances of clinical trials for CRC immunotherapy. The article will also discuss the reasons for the low efficacy of immunotherapeutic approaches in CRC and provide several potential directions for the future development of CRC immunotherapy.

Relationship between oncolytic viral therapy and tumor microenvironment or body immunity / 肿瘤研究与临床

The interaction between oncolytic virus (OV) and the tumor microenvironment or body immune system is critical to the outcome of antitumor therapy. The antitumor mechanism of OV is complex, which involves direct cytotoxic effects, immunogenicity change in tumor microenvironment, the role of tumor vasculature, and activating of the antitumor immunity response, to reach the goal of killing the tumor cells infected or uninfected, and confirming the continous favorable effects.

Progress in systemic administration of adenovirus vector for cancer gene therapy / 肿瘤

Gene therapy has become a new cancer therapy model behind surgical excision, radiotherapy, chemotherapy and interventional therapy. The choice of proper carrier is very important in cancer gene therapy. Adenovirus is widely used as a vector carrier in cancer gene therapy. Gene-engineered oncolytic adenovirus (OncoAd) has the advantages including cancer cell-specific replication, infected cell destruction, and high expression of inserted therapeutic genes, so as to obtain the potent antitumor efficacy. Therefore, using OncoAd is considered to be an effective tumor therapy strategy. The systemic administration of intravenous delivery plays an important role in the treatment of advanced metastatic cancer. However, the delivery of OncoAd was limited to local injection in vivo in the past. Because the efficacy of intravenously administering OncoAd is compromised by non-specific sequestration in the liver and host immune response, while the non-viral vectors have the advantages of good biosafety and low immunogenicity, the combination of the two will be beneficial to the systemic administration of OncoAd. In this paper, the research strategies of OncoAd intravenous delivery for cancer gene therapy in recent years are reviewed, and the research progress in OncoAd combined with non-viral vector for cancer gene therapy is emphasized.

Intratumor injection of recombined oncolytic adenovirus and ethanol in advanced pancreatic carcinoma combined with intravenous gemcitabine: a comparative study / 中华消化内镜杂志

Objective To compare the efficacy and safety of E1B gene-deleted adenovirus (H101)and ethanol in treating advanced pancreatic carcinomas by intratumoral injection combined with intravenous gemcitabine.Methods We constructed an orthotopic nude mouse model of pancreatic carcinoma through cancer cell injection into pancreas.A total of 54 nude mice were randomly allocated to 6 groups to accept H101,ethanol or saline (control) intratumoral injection,combined with or without intravenous gemcitabiein.The animals were sacrificed 4 weeks after the treatment and the pancreatic tumors were collected to determine the size,existence of metastasis,distribution of virus by indirect immunofluorescence and apoptosis in tumor by TUNEL and electron microscope.Results All mice completed the scheduled treatment,while 3 died in 48 hours after ethanol injection resulting in a mortality of 16.7％ (3/18).On the contrary,no mice died in the adenovirus injcction group.The average tumor size in group of H101 intratumoral injection combined with intravenous gemcitabie was significant smaller than that in group of saline injection with or without systemic gemcitabie (P =0.008,0.040,respectively).Similar differences were observed between ethanol intratumoral injection and control groups (P =0.012,0.041).Meanwhile,the H101 was absent in all the other organs except the pancreas,which meant that the selectivity of the H101 was tremcndous.The virus combine gemcitabie group had higher apoptosis rate in tumor (83.2 ± 35.7) ％,determined by TUNEL.Conclusion E1B gene-deleted adenovirus intratumral injection in combination with intravenous gemcitabine treating pancreatic carcinomas is efficient and safe,in spite of its lower effectiveness than ethanol.

Oncolytic viruses & their specific targeting to tumour cells.

Cancer is one of the major causes of death worldwide. In spite of achieving significant successes in medical sciences in the past few decades, the number of deaths due to cancer remains unchecked. The conventional chemotherapy and radiotherapy have limited therapeutic index and a plethora of treatment related side effects. This situation has provided an impetus for search of novel therapeutic strategies that can selectively destroy the tumour cells, leaving the normal cells unharmed. Viral oncotherapy is such a promising treatment modality that offers unique opportunity for tumour targeting. Numerous viruses with inherent anti-cancer activity have been identified and are in different phases of clinical trials. In the era of modern biotechnology and with better understanding of cancer biology and virology, it has become feasible to engineer the oncolytic viruses (OVs) to increase their tumour selectivity and enhance their oncolytic activity. In this review, the mechanisms by which oncolytic viruses kill the tumour cells have been discussed as also the development made in virotherapy for cancer treatment with emphasis on their tumour specific targeting.

Updates on current advances in gene therapy / Actualizaciones acerca de los adelantos presentes en terapia genética

Gene therapy is the attempt to treat diseases by means of genetic manipulation. Numerous challenges remain to be overcome before it becomes available as a safe and effective treatment option. Retroviruses and adenoviruses are among the most commonly used viral vectors in trials. The retrovirus introduces the gene it carries into the target cell genome while the adenovirus introduces the gene into the target cell nucleus without incorporating it into the target cell genome. Other viral vectors such as adenoassociated viruses, pseudotyped viruses and herpes simplex viruses, are also gaining popularity. Proposed nonviral methods for gene transfer include physical methods and the employment of chemical vectors (lipoplexes, polyplexes and inorganic nanoparticles). Recent studies have investigated potential applications of gene therapy in correcting genetic diseases, treating malignant disorders and for treatment of other diseases. Trials on gene therapy for SCID and Leber's congenital amaurosis have achieved considerable success, but the widely publicized adverse reaction in Xlinked SCID patient receiving gene therapy raised concerns for safety profile of gene therapy. For that, several methods of improving safety and efficacy of gene therapy have been proposed. At present, the three main gene therapy strategies for treatment of cancer are application to oncolytic viruses, suicidegene therapy and genebased immunotherapy. Gendicine, the first approved anticancer drugs based on the use of gene therapy principle, is based on the use of oncolytic viruses. More evidence for wider clinical applications of gene therapy are expected as more gene therapy studies progress from the preclinical phase to clinical trial.

La terapia genética es el intento de tratar enfermedades por medio de la manipulación genética. Quedan aún numerosos retos que superar antes de que esté tipo de tratamiento se encuentre disponible como una opción segura y eficaz. Los retrovirus y los adenovirus se hallan entre los vectores virales más comúnmente utilizados en ensayos: el retrovirus introduce el gen - del cual es portador - en el genoma de la célula de destino, mientras el adenovirus introduce el gen en el núcleo de la célula de destino sin incorporarlo al genoma de la célula de destino. Otros vectores virales tales como los virus adenoasociados, los virus pseudotipados, y los virus del herpe simple, también están ganando popularidad. Los métodos no virales propuestos para la transferencia de genes incluyen tanto métodos físicos como el empleo de vectores químicos (lipoplexes, polisomas y nanopartículas inorgánicas). Estudios recientes han investigado las aplicaciones potenciales de la terapia genética en la corrección de las enfermedades genéticas, el tratamiento de los trastornos malignos y para el tratamiento de otras enfermedades. Los ensayos de terapia genética para SCID y la amaurosis congénita de Leber han logrado un éxito considerable, pero la reacción adversa ampliamente divulgada en el caso de los pacientes con SCID ligada al cromosoma, que recibían terapia génica, causó preocupación en cuanto al perfil de seguridad de la terapia génica. Por esa razón, se han propuesto varios métodos para mejorar la seguridad y la eficacia de la terapia génica. En la actualidad, las tres estrategias principales de terapia de genes para el tratamiento del cáncer son la aplicación de virus oncolíticos, la terapia con gen suicida, y la inmunoterapia genética. La gendicina, el primer medicamento anticancerígeno aprobado, basado en el uso del principio de la terapia génica, se basa en el uso de virus oncolíticos. Se esperan más evidencias a favor de aplicaciones clínicas más amplias de la terapia génica, a medida que más estudios de terapia génica progresan de la fase preclínica a la fase de ensayo clínico.

The effect of oncolyic adenovirus SG600-IL24 expressing human MDA-7/IL-24 on apoptosis of hepatocellular carcinoma cell lines / 中华普通外科杂志

Objective To investigate the effect of oncolytic adenovirus vector SG600-IL24expressing human melanoma differentiation associated gene-7 (mda-7/IL-24) on hepatocellular carcinoma cell lines with different metastatic potential of HepG2, SMMC7721, MHCC97L and normal liver cell line LO2. Methods The oncolytic adenovirus SG600-IL24 which carrying mda-7/IL-24 gene was transfected into hepatocellular carcinoma cell lines and normal liver cell line. The mRNA and protein expression of mda7/IL-24 in HepG2, SMMC7721, MHCC97L and LO2 cell lines was confirmed by RT-PCR,ELISA assay and Western blot respectively. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst33258 and flow cytometry were studied to indicate the apoptosis effects. Results It was confirmed by RT-PCR, ELISA assay and Western-blot that the exogenous mda-7/IL-24 gene was highly expressed in HepG2, SMMC7721, MHCC97L and LO2 cell lines. MTT and apoptosis detection indicated that MDA-7/IL-24 can induce the growth suppression (the inhibition rate was 75% ±2. 5% ,86% ±3. 5% ,and promotes apoptosis ( the apoptosis rate was 56. 5% ± 4. 0% , 34. 4% ± 2. 0% , 43. 3% ± 2. 5%cell lines at G2/M phase ( the blocking rate was 35. 4% ± 4. 2% , 40. 5% ± 5. 0% , 42. 0% ± 5. 0%metastatic potential hepatocellular carcinoma cell lines but not in normal liver cell line.Conclusions Oncolytic adenovirus vector SG600-IL24 can selectively induce growth suppression, promote apoptosis in hepatocellular carcinoma lines in vitro but not in normal liver cell LO2.

Targeting oncolytic adenovirus therapy in pancreatic cancer / 国际肿瘤学杂志

Carrying out targeting an oncolytic adenovirus strategy in gene therapy of pancreatic cancer is a new direction.Currently,related research including vitro test and animal models vivo test,targeted strategies on molecular biology and gene level are carried out.Some of oncolytic adenovirus drugs have entered phase Ⅲ clinical trials.Oncolytic adenoviruses alone or in combination with other treatments,can enhanced anti-tumor effect.

Cell-based delivery of oncolytic viruses / 国际肿瘤学杂志

Oncolytic viruses,a novel class of virus vectors,which selectively replicate only in tumor cells,have excellent tumor targeting and good tansfection efficiency.Many oncolytic viruses have apparent curative effect when administered intratumorally.However,the host immune system remains a critical obstacle to systemic administration of virotherapeutics.It appears that cell-based delivery of oncolytic viruses could offer one solution to this critical problem,which provides a new platform to the biological therapy of cancer.

Oncolytic adenoviruses harboring IL-24 gene induce the apoptosis of melanoma cells / 中华皮肤科杂志

Objective To study the effects of oncolytic adenoviruses ZD55 harboring IL-24 gene (ZD55-IL-24) on the apoptosis of human melanoma cell line A375. Methods The oncolytie adenoviruses ZD55-IL-24 were verified by PCR. Then, the viruses were propagated, purified, and titrated by HEK293 cell plaque assay. A375 cells were cultured, divided into three groups transfected with ZD55-1L-24, ZD55 fused with enhanced green fluorescent protein (ZD55-EGFP), and replication-deficient adenovirus ZD55 carrying IL-24 gene (AD-IL-24), respectively. The multiplicity of infection was 0.1, 1, 10 and 100, respectively.Subsequently, the eytotoxity of these viruses and proliferation of A375 cells were determined by crystal violet staining and methyl thiazolyl tetrazolium (MTT) assay, respectively. The expressions of EIA and IL-24 protein were detected by Western blot in A375 cells. Results PCR verified that the adenoviruses ZD55-IL-24 contained IL-24 gene without wild adenovirns contamination. Crystal violet staining revealed that ZD55-IL-24 had an obvious eytotoxic effect on A375 cells, and MTT assay indicated that ZD55-IL-24 inhibited the proliferation of A375 cells in a time-and concentration-dependent manner. As shown by Western blot analysis, ZD55-1L-24 expressed IL-24 and E1A protein in A375 cells with a high efficiency. Conclusions The oncolytic adenoviruses ZD55-IL-24 can efficiently express IL-24 gene, inhibit the proliferation of, and induce the apoptosis in A375 cells.

Advancement of Reovirus-based therapy for cancer / 国际肿瘤学杂志

Reovirus,a kind of oncolytic viruses, is seldom pathogenic, but is selectively able to replicate in cancer cells through activation of Ras signaling. Pre-clinical studies have demonstrated that treatment with reovirus is associated with significant anticancer activity across a range of tumor types. Further clinical evaluation of reovirus therapy has shown that it is well tolerated when administered locally or systemically. Encouraging anticancer efficacy has been observed with single-agent treatment and in combination with chemotherapy and radiotherapy. High safety and promising efficacy of reovirus has raised hopes that it will become a new anticancqt agent.

Oncolytic adenovirus in tumor targeted therapy / 国际肿瘤学杂志

Oncolytic adenovirus is known to carry out the transformation of adenovirus,which has kill-ing role on tumor defective gene phenotypes,and Can be used as vector for tumor targeted tbempy drugs.At pres-ent,H101 and dl1520 and other biological products are gained from adenovirus type 5-specific gene fragments,which targeted tumor cell defects in the p53 gene expression,can specifically recognize tumor cells and Carry out proliferation and lysis of tumor cells.These biological products can be used as vectors to carry the anti-tumor genes and result in more powerful anti-tumor effects.

EUS-guided oncolytic adenovirus implantation in patients of non-operative pancreatic cancer / 中华消化内镜杂志

Objective To observe the safety and efficacy of oncolytic adenovirus (H101) implantation under EUS guidance combined with gemcitabine in patients of non-operative pancreatic cancer.Methods From May 2007 to December 2007,6 patients with non-operative pancreatic cancer were enrolled in the study.H101 were implanted into 3 sites of the tumor under EUS guidance.Gemcitabine Was siren systemicly on d2,d9 and d16 after implantation, and repeated 1 month later.Tumor size and perfusion were assessed by computed reconstruction and perfusion imaging.Changes of clinical indices,adverse effects and complications were also recorded.Results All patients completed the two periods of treatment as planned.Tumor size decreased in 5 cases(18.21%-38.65%),but without statistical difference (P=0.078).The area of liver metastasis Was found decreased in 1 ease.Perfusion imaging showed significant increase of mean transit time(P=0.049) and improvement in blood flow,blood volume and permeability surface at 2 weeks after the treatment. KPS increased in 2 patients and pain score decreased in 3 patients.Three patients died 2.5,2.5 and 3 months respectively after the procedure.while 3 other patients are still alive with the survival time of 3,5 and 10 months. Major adverse effects associated with H101 implantation were fever and flu-like symptoms.Mild acute pancreatitis occurred in 1 cage.Conclusion EUS guided oncolytic adenovirus implantation in advanced pancreatic cancer is feasible and safe.With the combination of gemcitabine,it is capable of shrinking the tumor size,destroying the angiogenesis of the tumor and improving the patients'living quality.

In vitro study on combination of oncolytic adenovirus and Ad - Endo against cervical carcinoma / 中国基层医药

Objective To evaluate the anti-cervical carcinoma cells effects of oncolytic adenovirus d/1520 and Ad-Endo in vitro,and investigate the influence of d/1520 on the Endostatin expression directed by Ad-Endo in cervical carcinoma cells.Methods After cervical carcinoma cells were infected with d/1520 or/and Ad-Endo,the cytopathic effects (CPE) were observed,and the cytotoxicities were measured by MTT assay.Endestatin concentrations were measured by ELISA.Results d/1520 can replicate in cervical carcinoma cells and induce cytopathie effects; In MTT assay,Ad-Endo hardly inhibited the growth of cervical carcinoma cells,while d/1520 inhibited efficiently the proliferation of cervical cancer cells.And the inhibitory rates in Ad-Endo + d/1520 group were higher than that in d/1520 group.Ad-Endo expressed Endestatin protein in cervical cancer cells,while Endostatin concentrations in Ad-Endo+dl1520 group were much higer than those in Ad-Endo group.Conclusions dl1520 can kill cervical carcinoma cells in vitro,and this eytotoxieity can be enhanced by combining with Ad-Endo.The expression of Endostatin directed by Ad-Endo in cervical cancer cells can be dramatically increased by dl1520.