Expressions of HBXIP and GRIM-19 in hepatocellular carcinoma and their clinic significance / 天津医药

Objective To investigate the expressions of HBXIP and GRIM-19 in hepatocellular carcinoma tissues and their clinic significance. Methods Hepatocellular carcinoma tissue (n=42) and normal liver tissue (n=28) were collected from Tianjin First Central Hospital,immunohistochemistry was used to detect the expressions of HBXIP and GRIM-19 in these two groups. Results Rate of cells with positive expressions of HBXIP in hepatocellular carcinoma and normal liver tissues were 80.95%(34/42)and 42.86%(12/28)respectively;Rate of cells with positive expression of GRIM-19 in hepato?cellular carcinoma tissues and normal liver tissues was 40.48%(17/42)and 75.00%(21/28)respectively, and the difference between these two groups was statistically significant(P<0.05);The expression of HBXIP was higher but the expression of GRIM-19 was lower in poor differentiated and stageⅢ-IV cells than those in well and moderate differentiated cells and in stage I-II, cells. What′s more, the expression of GRIM-19 is higher in tissue without portal thrombosis than that in tissue with portal thrombosis. The expression of HBXIP was negatively correlated with GRIM-19 expression(rS=-0.400,P<0.01). Conclusion The abnormal expressions of HBXIP and GRIM-19 may play important roles in the process of development and metastasis of hepatocellular carcinoma.

Research advancement of pro-apoptotic mechanisms of GRIM-19 and its related proteins / 医学研究生学报

Cancer development is a complex process that involves multiple genetic changes and multiple signaling pathways . Recent findings show that the GRIM-19 is a novel apoptosis regulation gene , and its gene mutations and loss of protein expression have been observed in many tumor types such as urinarysystem tumor , digestive system neoplasm , which are closely related to cancer devel-opment.Thus, GRIM-19 may be a potential target for gene therapy .Pro-apoptotic mechanisms of GRIM-19 and its related proteins such as STAT3,GW112,p16INK4aare overviewed in this article.

Attenuated Salmonella carrying pGRIM-19-si-survivin co-expression plas-mid inhibits growth of prostate cancer subcutaneous xenografts in vivo / 中国病理生理杂志

AIM:To explore the effects of pGRIM-19-si-survivin co-expression plasmid carried by human at-tenuated Salmonella on prostate cancer subcutaneous xenograft growth in nude mice .METHODS: Prostate cancer xeno-graft model was established in nude mice .Co-expression plasmids carried by attenuated Salmonella were introduced by in-traperitoneal injection .The xenograft volumes were monitored timely .Immunohistochemical staining , RT-PCR and TUNEL assay were applied to investigate the related mechanisms that pGRIM-19-si-survivin inhibited tumor growth in vivo.RE-SULTS:Compared with psi-survivin and pGRIM-19 carried by attenuated Salmonella ( control groups ) , the tumor volumes were reduced markedly in pGRIM-19-si-survivin plasmid group .The mean shrinkage rates were 2.36 and 3.02 times. pGRIM-19-si-survivin co-expression plasmid carried by attenuated Salmonella inhibited survivin expression but strengthened GRIM-19 expression obviously (P<0.05).The mRNA expression of apoptosis-related proteins such as Bcl-xL, Stat3, cy-clin D1 and c-Myc was inhibited , and the vascular endothelial growth factor ( VEGF) mRNA and Ki67 protein were also in-hibited, but the caspase-3 mRNA expression was up-regulated ( P<0.05 ) with significant cell apoptosis .CONCLU-SION:pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella inhibits the growth of prostate cancer subcutaneous xenografts in nude mice by promoting cell apoptosis and inhibiting prostatic cancer proliferation .

Expression of GRIM-19 and its target gene product STAT3 in human breast cancer / 中华内分泌外科杂志

Objective To evaluate gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) expression levels and the correlation with its target gene product signal transducers and activator of transcription 3 (STAT3)in human breast cancer tissues and normal gland tissues,and to analyze their roles in the tumorigenesis of breast cancer.Methods The expression of GRIM-19 and STAT3 protein and mRNA in 40 cases of breast cancer tissues and 40 cases of normal gland tissues was detected by immunohistochemistry and Western blot.The correlation of the expression of GRIM-19 and STAT3 to various clinicopathologic characteristics of breast cancer were analyzed statistically.The mRNA expression and gene mutation of GRIM-19 in breast cancer cell line MCF-7 and 25 specimens of breast cancer and normal gland tissue were detected by reverse transcription-polymerase chain reaction(RT-PCR)and sequencing.Results The protein and mRNA expression of GRIM-19 was obviously lower in breast cancer than in normal gland tissues (P ＜ 0.05) while the protein and mRNA expression of STAT3 was obviously higher in breast cancer than in normal gland tissues(P ＜0.05).The expression of GRIM-19 and STAT3 was negatively correlated with each other(x2 =8.25,P ＜0.01).Breast cancer samples exhibited low level of GRIM-19 and moderate to high level of STAT3 expression.In contrast,the normal gland tissue was characterized by high level of GRIP-19 and low level of STAT3 expression.The protein expression of GRIM-19 was correlated with the histological grading and clinical stage of breast cancer(P ＜ 0.05).STAT3 was not correlated with clinicopathologic characteristics of breast cancer (P ＞ 0.05).No mutation of GRIM-19 gene was detected in breast cancer tissues,normal gland tissues or MCF-7 breastcancer cells.Conclusions The low expression of GRIM-19 and the high expression of STAT3 co-exist in breast cancer.Downregulation of GRIM-19 was closely correlated with increased histological grade,clinical stage and STAT3 in breast cancer.The potential role of GRIM-19 in breast cancer development may be through these correlations.

Relationship between GRIM-19 and cancers / 国际外科学杂志

Gene associated with retinoid-IFN-induced mortality-19(GRIM-19)is originally isolated as a growth suppressive gene using a genetic screen.GRIM-19, initially found in nucleus and mitochondria, is essential for the assembly and functioning of mitochondrial complex I.GRIM-19 involves the regulation of cell proliferation and apoptosis, and low expression or mutation of GRIM-19 contributes to abnormal proliferation.During viral oncogenesis, GRIM-19 may be a general target protein similar to other cellular tumor suppressors.GRIM-19 plays an important role in tumor formation and apoptosis inhibition, and may be as a new tumor marker to early cancer screening.

Grim-19 overepression induces apoptosis of colon cancer SW480 cells / 中国肿瘤生物治疗杂志

Objective:To investigate the effect of retinoid-interferon-induced mortality (GRIM-19) gene on the apoptosis of colon cancer. Methods: A GRIM-19 eukaryotic expression vector (pCMV-Flag-GRIM-19) was constructed and transfected into SW480 cells. Expressions of GRIM-19 and apoptosis-related proteins were detected by Western blotting analysis. Apoptosis of SW480 cells was measured by Annexin-V/PI assay and mitochondrial membrane potential JC-1 staining. Results: The GRIM-19 eukaryotic expression vector pCMV-Flag-GRIM-19 was successfully constructed. Expression of GRIM-19 in SW480 cells was up-regulated and that of apoptosis-related protein Bcl-xl was down-regulated after transfection with pCMV-Flag-GRIM-19. Apoptosis rate was (7.7±1.39)% in SW480 cells transfected with pCMV-Flag empty vector and (15.0 ± 2.52)% in pCMV-Flag-GRIM-19 transfected cells (P<0.05). Mitochondrial membrane potential was decreased in (7.5±2.09)% of pCMV-Flag transfected cells and (17.5±3.07)% of pCMV-Flag-GRIM-19 transfected cells (P<0.05). Conclusion: In vitro GRIM-19 transfection can effectively induce apoptosis of colon cancer SW480 cells.

GRIM-19 Expression and Function in Human Gliomas

OBJECTIVE: We determined whether the expression of GRIM-19 is correlated with pathologic types and malignant grades in gliomas, and determined the function of GRIM-19 in human gliomas. METHODS: Tumor tissues were isolated and frozen at -80degrees C just after surgery. The tissues consisted of normal brain tissue (4), astrocytomas (2), anaplastic astrocytomas (2), oligodendrogliomas (13), anaplastic oligodendrogliomas (11), and glioblastomas (16). To profile tumor-related genes, we applied RNA differential display using a Genefishing(TM) DEG kit, and validated the tumor-related genes by reverse transcription polymerase chain reaction (RT-PCR). A human glioblastoma cell line (U343MG-A) was used for the GRIM-19 functional studies. The morphologic and cytoskeletal changes were examined via light and confocal microscopy. The migratory and invasive abilities were investigated by the simple scratch technique and Matrigel assay. The antiproliferative activity was determined by thiazolyl blue Tetrazolium bromide (MTT) assay and FACS analysis. RESULTS: Based on RT-PCR analysis, the expression of GRIM-19 was higher in astrocytic tumors than oligodendroglial tumors. The expression of GRIM-19 was higher in high-grade tumors than low-grade tumors or normal brain tissue; glioblastomas showed the highest expression. After transfection of GRIM-19 into U343MG-A, the morphology of the sense-transfection cells became larger and more spindly. The antisense-transfection cells became smaller and rounder compared with wild type U343MG-A. The MTT assay showed that the sense-transfection cells were more sensitive to the combination of interferon-beta and retinoic acid than U343MG-A cells or antisense-transfection cells; the anti-proliferative activity was related to apoptosis. CONCLUSION: GRIM-19 may be one of the gene profiles which regulate cell death via apoptosis in human gliomas.

Construction of Recombinant Adenovirus Carrying GRIM19 and Its Effect on SW480 Cells / 华中科技大学学报（医学）（英德文版）

In order to examine the effect of GRIM 19 on colon cancer cell SW480, the recombinant adenovirus carrying GRIM19 gene was constructed and transfected into SW480 cells. GRIMI9 cDNA was amplified by PCR with the template pcxn2-GRlMl9 and cloned into the shuttle plasmid pAdTrack-CMV. The plasmid pAdTrack-CMV-GRIM19 was linearized by PmeI and homologously recombined with bone plasmid pAdEasy-1 in BJ5183, followed by identification by enzyme diges- tion. After transfection of linearized pAd-GRIM19 with PacI into HEK293 cells, Ad-GRIMI9 was obtained and amplified by 3 circles. SW480 cells were infected with Ad-GRIM19. The apoptosis rate was detected by flow cytometry. Agarose electrophoresis revealed the bands of recombinant plasmids identified by enzyme digestion were in the right range corresponding with expectation. Under the fluorescent microscopy, the package of Ad-GRIM19 in HEK293 cells and the expression of Ad-GRIM19 in SW480 cells were observed. The transfection of Ad-GRIM19 into SW480 cells in-creased the apoptosis rate of SW480 cells as compared with controls. It was concluded that Ad-GRIM19 was successfully constructed and the overexpression of GRIM19 in colon cancer cell lines could promote the apoptotic cell death.