Expression change and role of myeloma cancer gene mRNA, microRNA-540-3p, circular RNA-04996 in the liver inflammatory reaction during the rat liver regeneration / 解剖学报

Objective To explore the role pathway and pattern of the transcription factor myeloma cancer gene (MYC) and its mRNA interaction with microRNA(miRNA, miR) and ciccular RNA(circRNAs) at 0 hour and 6 hour in the rat liver regeneration. Methods The rat 2/3 hepatectomy (partial hepatectomy，PH) model was prepared as described by Higgins, the expression changes of mRNA, miRNA and circRNA together named as competing endogenous RNAs(ceRNA) of remnant liver were detected by the large-scale quantitative detection technology, the interaction network of ceRNA was constructed by Cytoscape 3.2 software, and their correlation in expression and role were analyzed by ceRNA comprehensive analysis. Results It was found that at 0 hour and 6 hours after PH, the ratio value of MYC mRNA showed 0.15±0.03 and 2.36±0.20, miR-540-3p displays 3.00±0.43 and 0.79±0.01, circRNA_04996 showed 1.43±0.43 and 3.14±0.94. At the same time, the four kinds of inflammatory reaction-related genes plasminogen activator urokinase receptor (PLAUR), tumor necrosis factor (TNF), interleukin 1 receptor type 2 (IL1R2), ect, which were prometed in expression by MYC, were down-regulated at 0 hour after PH, but the inflammatory reaction-related genes natriuretic peptide A (NPPA), nuclear receptor subfamily O group B member 2 (NROB2) and peroxisome proliferator activated receptor alpha (PPARA), which were inhibited in expression by MYC, were up-regulated at 0 hour after PH. On the other hand, the three kinds of inflammatory reaction-related genes PLAUR, TNF, IL1R2, ect, which are prometed in expression by MYC, were up-regulated at 6 hours after PH, but the inflammatory reaction-related genes NPPA, NROB2 and PPARA, which were inhibited in expression by MYC, were down-regulated at 6 hours after PH. Conclusion The correlation in expression and role of the miRNA, which are inhibited by circRNAs, MYC, its mRNA is inhibited by miRNAs, and the inflammatory reaction-related genes, which are regulated by MYC, and are helpful for the hepatocyte to be in non-inflammatory reaction state at 0 hour after PH and to be in inflammatory reaction state at 6 hours after PH.

An epidermal growth factor motif of developmental endothelial locus 1 protein inhibits efficient angiogenesis in explanted squamous cell carcinoma in vivo

ABSTRACT Background: Cancer gene therapy using a nonviral vector is expected to be repeatable, safe, and inexpensive, and to have long-term effectiveness. Gene therapy using the E3 and C1 (E3C1) domain of developmental endothelial locus-1 (Del1) has been shown to improve prognosis in a mouse transplanted tumor model. Objective: In this study, we examined how this treatment affects angiogenesis in mouse transplanted tumors. Materials and methods: Mouse transplanted tumors (SCCKN human squamous carcinoma cell line) were injected locally with a nonviral plasmid vector encoding E3C1 weekly. Histochemical analysis of the transplanted tumors was then performed to assess the effects of E3C1 on prognosis. Results: All mice in the control group had died or reached an endpoint within 39 days. In contrast, one of ten mice in the E3C1 group had died by day 39, and eight of ten had died or reached an endpoint by day 120 (p < 0.01). Enhanced apoptosis in tumor stroma was seen on histochemical analyses, as was inhibited tumor angiogenesis in E3C1-treated mice. In addition, western blot analysis showed decreases in active Notch and HEY1 proteins. Conclusion: These findings indicate that cancer gene therapy using a nonviral vector encoding E3C1 significantly improved life-span by inhibiting tumor angiogenesis. (REV INVEST CLIN. 2021;73(1):39-51)

Housekeeping genes involved in non-malignant breast phenotypes are widely expressed in multiple cancers and provide novel biomarkers of tumor classification

Clinically relevant biomarkers are useful to determine cancer patients' prognosis and treatments. To discover new putative biomarkers, we performed in silico analysis of a 325-gene panel previously associated with breast epithelial cell biology and clinical outcomes. Sixteen public datasets of microarray samples representing 8 cancer types and a total of 3,663 patients' samples were used for the analyses. Feature selection was used to identify the best subsets of the 325 genes for each classification, and linear discriminant analysis was used to quantify the accuracy of the classifications. A subset of 102 of the 325 genes were found to be housekeeping (HK) genes, and the classifications were repeated using only the 102 HK subset. The 325-gene panel and 102 HK subset were able to distinguish colon, gastric, lung, ovarian, pancreatic, and prostate tumors and leukemia from normal adjacent tissue, and classify disease subtypes of breast and lung cancers and leukemia with 70% or higher accuracy. HK genes have been overlooked as potential biomarkers due to their relative stability. This study describes a set of HK genes as putative biomarkers applicable to multiple cancer types worth following in subsequent validation studies.

Importance of MACC1 expression in breast cancer and its relationship with pathological prognostic markers

Background: Metastasis associated colon cancer gene 1 (MACC1) is a gene that was first described as a c-Met transcription regulator causing the progression of colon cancer. In this study, protein and messenger RNA (mRNA) expression of MACC1 in breast cancer and its relationship with clinicopathological prognostic parameters were investigated. Methods: Sixty-six cases with tumors underwent radical mastectomy for invasive ductal carcinoma and 25 control cases operated for mammoplasty were included in the study. In paraffin blocks of tumor and control tissues, MACC1 expression was investigated by the immunohistochemical method and Real-time polymerase chain reaction (Real-Time PCR). In addition, vascular endothelial growth factor (VEGF) expression was examined immunohistochemically in tumor tissues. The relationship between MACC1 expression in tumor tissues, clinicopathological prognostic parameters, and VEGF was investigated. Results: In this study, protein and mRNA expressions of MACC1 were found to be higher in tumor tissues compared with normal breast tissues. MACC1 protein expression was also associated with significant poor prognostic markers, such as high histologic grade, ER negativity, and HER2 positivity. However, there was no correlation between MACC1 expression and VEGF. Conclusion: According to these results, MACC1 expression may be a marker of breast carcinoma as well as an independent predictor of poor prognosis. In addition, MACC1 may not affect angiogenesis in breast cancer or even if it has an effect, it may not be associated with VEGF. However, it would be appropriate to support these results in a larger series by investigating in vivo and in vitro studies.

Implication of BRCA1/2 gene detection in malignant tumors / 中国肿瘤临床

Tumor suppressor genes breast cancer gene 1/2 ( BRCA1/2) play key roles in DNA damage repair pathways and are essential for maintaining genome integrity. The earliest studies found that BRCA1/2 are the main pathogenic factors of hereditary breast/ovari-an cancer syndrome, and the risk of breast cancer and ovarian cancer is greatly increased in BRCA1/2 mutation carriers. Recent studies have demonstrated that BRCA1/2 mutations also increased the risk of developing colon cancer, pancreatic cancer, skin cancer, and male prostate cancer. BRCA1/2 mutation patients have common molecular pathological basis, which may be independent of pathologi-cal tissue diagnosis in the future. Furthermore, it can serve as an important basis for clinical drug therapy, including chemotherapy drugs represented by platinum, PARP inhibitors, PD-1 antibodies, ALDH2 inhibitors, and mTOR inhibitors.

Applicable Value of AMSS-PCR in Lung Cancer Gene Mutation Detection / 中国肺癌杂志

BACKGROUND@#The detection of driver oncogenes of lung cancer is of great importance. There are various gene detection techniques nowadays which are different from each other. We carried out this study to investigate the specificity and sensitivity of assay panels based on an Amplification Refractory Mutation System-polymerase chain reaction (ARMS-PCR) technique of Amplification Mutation Specific System (AMSS) in detection of lung cancer gene mutation. To estimate the applicable value of assay panels in clinical settings.@*METHODS@#We collected cancer tissue specimens or fluid specimens from 309 patients. Mutation results were presented for those samples previously detected by ARMS-PCR. In comparison, we carried out AMSS-PCR using (epidermal growth factor receptor, EGFR) assay panel and Six-Alliance assay panel as well as Sanger sequencing. Software SPSS 22.0 (SPSS IBM) was used for statistical analysis.@*RESULTS@#The rates of consistency between the results by assay panels and Sanger sequencing or ARMS-PCR were 97.41% and 97.73%, respectively. Besides, EGFR assay panel had higher consistency rates with other detection methods than Six-Alliance assay panel. As for consistency test, the Kappa values of assay panels with Sanger sequencing, assay panels with ARMS-PCR, and ARMS-PCR with Sanger sequencing were 0.946, 0.953, and 0.913, respectively. The receiver operating characteristic curve (ROC) area under curve (AUC) of assay panels was 0.976 referring to Sanger sequencing, and 0.975 as ARMS-PCR was referred to.@*CONCLUSIONS@#AMSS-PCR can make an optimal cancer gene mutation detection method for clinical settings.

Applied research of prostate specific antigen combined with prostate cancer gene 3 in diagnosis of prostate disease / 中华内分泌外科杂志

Objective To detect the expression of peripheral blood free prostate specific antigen (fSPA),total prostate specific antigen (tSPA),prostate specific antigen density (PSAD) and prostate cancer gene 3 (PCA3) in prostate disease,and the significance of combined detection of fSPA,tSPA and PCA3.Methods 67 patients with prostate cancer,75 patients with prostatic hyperplasia and 70 healthy male were selected as the research objects from Dec.2014 to Jul.2016.The serum level of fSPA and tSPA was detected by chemiluminescence immune staining method.The prostate volume was tested by ultrasonic sound and PSAD value was calculated.The total RNA was extracted by Trizol,and the serum PCA3 mRNA expression was detected by RT-PCR.The specificity and sensitivity of combined detection of fSPA,tSPA and PCA3 were analyzed.Results The serum levels of fSPA,tSPA,PSAD and PCA3 in prostate cancer were significantly higher than those in patients with prostatic hyperplasia and healthy male,and they were higher in patients with hyperplasia of prostate than in healthy male,and the differences had statistical significance (P＜0.01).The serum levels of fSPA,tSPA,PSAD and PCA3 were higher in patients with Gleason score ≥7 points and in T3-T4 stage than in patients with Gleason score ＜7 and in T1-T2 stage,and the difference had statistical significance (P＜0.01).The serum levels of fSPA,tSPA,PSAD and PCA3 were positively correlated with Gleason score and TMN pathological stage,and the difference had statistical significance (P＜0.01).The AUC value of fSPA,tSPA,PSAD and PCA3 in diagnosis of prostate cancer was 0.53,0.57,0.63 and 0.75,and the AUC value of combined detection was 0.92.The combined detection efficiency was higher than the single index.The specificity of fSPA,tSPA,PSAD and PCA3 was 67.16％,68.66％,73.13％ and 85.07％,and the sensitivity was 71.64％,70.15％,74.63％ and 82.09％ respectively.The specificity of combined detection was 97.01％,the sensitivity was 92.54％,and the difference had statistical significance (P＜0.01).Conclusion The serum level of fSPA,tSPA,PSAD and PCA3 is increased in prostate disease,and is negatively correlated with Gleason score and TMN pathological stage.The combined detection of fSPA,tSPA,PSAD and PCA3 can improve the sensitivity and specificity of prostate disease diagnosis,and is of high clinical value.

Expression and significance of hypermethylation in cancer 1 protein and ovarian cancer gene 1 protein in ovarian cancer tissue / 重庆医学

Objective To study the expression of hypermethylated gene 1 protein and ovarian cancer gene 1 protein in ovarian cancer and its relationship with the pathological features of ovarian cancer,and to explore its significance in ovarian cancer.Methods Sixty-three cases of ovarian cancer specimens and 63 cases of normal ovarian tissue were taken from January 2014 to December 2015 at the First Hospital of Jilin University.Western blot was used to detect the expression of hypermethylated gene 1 protein and ovarian cancer gene 1 protein in ovarian cancer tissues and normal ovarian tissues,and to analyze the relationship between the expression of two proteins and ovarian cancer.Results The expression of hypermethylated gene 1 protein and ovarian cancer gene 1 protein in ovarian cancer tissues were significantly lower than that in normal ovarian tissues (P＜0.05).There was no significant difference in the expression of hypermethylated gene 1 protein in different staging,different degree of differentiation and different pathological types in ovarian cancer (P＞0.05).The expression of ovarian cancer gene 1 protein in ovarian cancer stage Ⅰ was higher than that in stage Ⅱ and Ⅲ-Ⅳ (P＜0.05).The expression of ovarian cancer gene 1 protein in ovarian cancer stage Ⅱ was higher than that in stage Ⅲ-Ⅳ (P＜0.05).The expression of ovarian cancer gene 1 protein in high differentiation of ovarian cancer was significantly higher than that in moderately differentiated and poorly differentiated (P＜0.05).There was no significant difference between ovarian cancer gene 1 protein expression and poorly differentiated ovarian cancer (P＞0.05).There was no sig nificant difference in the expression of ovarian cancer gene 1 protein in different pathological types of ovarian cancer (P＞0.05).Conclusion Hypermethylatel gene,protein participate in the occurrence of ovarian cancer,the ovarian cancer gene 1 protein is only related to ovarian cancer clinical stage and degree of differentiation.

The value of urine derived prostate cancer gene 3 score for predicting the result of prostate biopsy : a meta-analysis / 中华泌尿外科杂志

Objective To systematic review the clinical value of urine derived prostate cancer gene 3 (PCA3) scores for predicting the result of prostate biopsy during diagnosing the prostate cancer.Methods Articles related to the value of urine derived PCA3 score for forecasting the result of prostate biopsy during diagnosing the prostate cancer,published from inception to November 2013,were retrieved in the databanks including PubMed (1966-2013),CNKI (1979-2013),EMBASE(1988-2013),and Cochrane liberary.Meta-Disc 1.4 software was used to analyze data.According to the inclusion and exclusion criteria,the literatures were screened.And the quality of included literatures were evaluated.The pooled sensitivity,specificity,the likelihood ratio(LR) were calculated and summary receiver operating characteristic curve(SROC curve) was made.Results A total of 22 articles with 3 060 prostate cancer cases and 5 307 normal or BPH cases were included.Since there was heterogeneity between the studies,the data was calculated by the random effect model.The pooled sensitivity of PCA3 score was 63％ (95％CI:0.61-0.65) and the pooled specificity was 70％ (95％CI:0.69-0.72),respectively.The positive (P-LR) and negative likelihood ratio (N-LR) were 2.38 (95％CI:2.05-2.77) and 0.51 (95％CI:0.46-0.58),respectively.The diagnosis odds ratio (DOR) was 4.88 (95％ CI:3.83-6.21).Summary receiver operating characteristic curve area under curve (SROC AUC) was 0.7445,Q* index was 0.689,respectively.Conclusions The performance of urine derived PCA3 score has higher accuracy of detection than other method in the diagnosis of prostate cancer.It can be a novel and optional non-invasive test and an important adjunct test for predicting the result of prostate biopsy.

MicroRNA in carcinogenesis & cancer diagnostics: A new paradigm.

MicroRNAs (miRNAs) are small 22-25 nucleotides long non-coding RNAs, that are conserved during evolution, and control gene expression in metazoan animals, plants, viruses, and bacteria primarily at post-transcriptional and transcriptional levels. MiRNAs ultimately regulate target gene expression by degrading the corresponding mRNA and/or inhibiting their translation. Currently, the critical functions of miRNAs have been established in regulating immune system, cell proliferation, differentiation and development, cancer and cell cycle by as yet unknown control mechanism. MiRNAs play an essential role in malignancy, and as tumour suppressors and oncogenes. Thus, discovery of new miRNAs will probably change the landscape of cancer genetics. Significantly different miRNA profiles can be assigned to various types of tumours, which could serve as phenotypic signatures for different cancers for their exploitation in cancer diagnostics, prognostics and therapeutics. If miRNA profiles can accurately predict malignancies, this technology could be exploited as a tool to surmount the diagnostic challenges. This review provides comprehensive and systematic information on miRNA biogenesis and their implications in human health.

Application value of cancer gene c-erbB-2 and its protein product expression in evaluation of the diagnosis and prognosis of breast cancer with lymph node metastasis / 中国基层医药

Objective To discuss the application value of cancer gene c-erbB-2 and its protein products P185 in evaluation of the diagnosis and prognosis of breast cancer with lymph node metastasis,and to explore the relationship between c-erbB-2 expression and tumor pathology classification,relapse.Methods 112 cases with mammary disease were selected as the research subjects.According to different clinical diagnosis,they were divided into breast cancer lymph node metastasis group,breast cancer without metastasis group and mammary gland benign pathological change group.The c-erbB-2 gene and carcinoembryonic antigen(CEA) were detected by immunohistochemical method,enzyme-linked immunosorbent(ELISA) and electrochemical luminescence method.The c-erbB-2 protein(P185)and CEA of the three groups were compared,the sensitivity and specificity of c-erbB-2 expression and CEA in the diagnosis of breast cancer with lymph node metastasis were analyzed.At the same time,the relationship between cerbB-2 expression and tumor pathology classification,relapse was analyzed.Results P185 and CEA index of breast cancer lymph node metastasis group and breast cancer without metastasis group were significantly higher than those of mammary gland benign pathological change group(t =7.093,6.443,9.232,8.508,all P ＜ 0.0 1),and P185 index of breast cancer lymph node metastasis group was significantly higher than without metastasis group (t =2.281,P ＜0.05),and there was no significant difference of CEA between the two breast cancer groups (t =0.406,P ＞ 0.05).The sensitivity of c-erbB-2 expression in dignosis of breast cancer lymph node metastasis had no difference with CEA (x2 =0.632,P ＞ 0.05),but the specificity was significantly higher than CEA(x2 =11.900,P ＜ 0.01).The positive expression rate of c-erbB-2 and tumor pathology classification had no obvious relation:Postoperative recurrence rate of c-erbB-2 positive expression patients was significantly higher than the negative expression patients(x2 =5.141,P ＜0.05).Conclusion c-erbB-2 gene and its protein product has good sensitivity and specificity in the diagnosis of breast cancer lymph node metastasis,and has obvious correlation with postoperative breast cancer recurrence,has important reference value in evaluation of the diagnosis and prognosis of breast cancer.

CaGe: A Web-Based Cancer Gene Annotation System for Cancer Genomics

High-throughput genomic technologies (HGTs), including next-generation DNA sequencing (NGS), microarray, and serial analysis of gene expression (SAGE), have become effective experimental tools for cancer genomics to identify cancer-associated somatic genomic alterations and genes. The main hurdle in cancer genomics is to identify the real causative mutations or genes out of many candidates from an HGT-based cancer genomic analysis. One useful approach is to refer to known cancer genes and associated information. The list of known cancer genes can be used to determine candidates of cancer driver mutations, while cancer gene-related information, including gene expression, protein-protein interaction, and pathways, can be useful for scoring novel candidates. Some cancer gene or mutation databases exist for this purpose, but few specialized tools exist for an automated analysis of a long gene list from an HGT-based cancer genomic analysis. This report presents a new web-accessible bioinformatic tool, called CaGe, a cancer genome annotation system for the assessment of candidates of cancer genes from HGT-based cancer genomics. The tool provides users with information on cancer-related genes, mutations, pathways, and associated annotations through annotation and browsing functions. With this tool, researchers can classify their candidate genes from cancer genome studies into either previously reported or novel categories of cancer genes and gain insight into underlying carcinogenic mechanisms through a pathway analysis. We show the usefulness of CaGe by assessing its performance in annotating somatic mutations from a published small cell lung cancer study.

The anti-tumor effect by adenovirus-mediated ING4 and IL-24 co-expression on hepatocellular carcinoma in vitro / 中华微生物学和免疫学杂志

Objective To construct a recombinant adenoviral vector carrying and co-expressing human inhibitor of growth 4(ING4) and human interleukin-24(IL-24) mediated by poly( A)-Promoter[Ad-ING4-poly(A)-Promoter-IL-24, referred to as Ad-ING4-IL-24] and explore its effect on the growth of HepG-2 human hepatocellular carcinoma cellsin vitro. Methods The poly(A)-Promoter(hEFl-elF4g) (Sal Ⅰ and Not Ⅰ ), ING4 ( Bgl Ⅱ and Sal Ⅰ ), and IL-24 ( Xho Ⅰ and Xba Ⅰ ) fragments were amplified by PCRusing pORF-mbcl-2α, pcDNA3.0-IL-24, and pcDNA3.0-ING4 plasmids as templates and subcloned into pAdTrack-CMV transfer vector to form pAdTrack-CMV-ING4-poly (A)-Promoter-lL-24, respectively. The pAdTrack-CMV-ING4-poly (A)-Promoter-IL-24 transfer vector linearized with Pme Ⅰ digestion and pAdEasy-1 backbone vector was further cotransformed into the bacteria BJ5183 competent cells for homologous recombination. The resultant pAdEasy-l-pAdTrack-CMV-ING4-poly ( A )-Promoter-IL-24 homologous recombinant plasmids were linearized with Pac Ⅰ digestion and transfected into the human embryonic kidney 293 (QBI-293A) cells by liposome, leading to formation of the recombinant adenoviruses Ad-ING4-IL-24co-expressing ING4 and IL-24. The Ad-ING4-IL-24 were amplified in QBI-293A cells and its titer was up to 3.5 × 109 PFU/ml. Adenovirus-mediated ING4 and IL-24 genes expression in HepG-2 cells was examined by RT-PCR and Western blot. The growth-suppressing and apoptosis-inducingg effect of Ad-ING4-IL-24 coexpressing ING4 and IL-24 on HepG-2 human hepatocellular carcinoma cells was assessed by MTT assay and FCM, respectively. Results DNA sequencing showed that the ING4, poly (A)-Promoter, and IL-24 fragments subcloned into pAdTrack-CMV plasmids were completely identical to those reported in GenBank.ING4 and IL-24 gene mediated by adenovirus could both successfully express in HepG-2 cells. Adenovirusmediated ING4 and IL-24 co-expression significantly suppressed HepG-2 hepatocellular carcinoma cell growth and induced cell apoptosis, and the effect of Ad-ING4-IL-24 group was more significant than AdING4 and Ad-IL-24 group. Conclusion The adenoviral vector co-expressing ING4 and IL-24 mediated by poly(A)-Promoter(Ad-ING4-IL-24) was successfully constructed. Ad-ING4-IL-24 had marked anti-tumor effect in suppressing HepG-2 human hepatocellular carcinoma cell growth and inducing cell apoptosis in vitro. Compared with Ad-ING4 and Ad- IL-24, Ad-ING4-IL-24 enhanced anti-tumor effect.

Replication-incompetent Adenovirus Vector-mediated MDA-7/IL-24 Selectively Induces Growth Suppression and Apoptosis of Hepatoma Cell Line SMMC-7721 / 华中科技大学学报（医学）（英德文版）

In order to investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 and normal liver cell line L02, the recombinant replication-incompetent Ad.mda-7 virus vector was constructed and infected into the HCC cell line SMMC-7721 and normal liver cell line L02. RT-PCR was performed to examine the expression of MDA-7 mRNA. The concentrations of MDA-7/IL-4 in culture supernatants were determined by using ELISA. MTT and Hoechst staining assay were applied to observe the inhibitory and killing effects of MDA-7 on the HCC cells. By using flow cytometry, the apoptosis, cell cycle and proliferation of SMMC-7721 and L02 cells were meas- ured. The results showed recombinant replication-incompetent virus expressing MDA-7/IL-24 was constructed successfully, and RT-PCR revealed that it could mediate the high expression of the ex- ogenous gene MDA-7/IL-24 in SMMC-7721 and L02 cells. The expression of MDA-7/IL24 proteins in the culture supernatant was detectable by ELISA. Ad.mda-7 infection induced apoptosis and growth suppression in SMMC-7721 cells and an increased percentage of HCC cells in the G2/M phase of the cell cycle, but not in L02 cells. It was concluded that mda-7/IL-24 gene, mediated with replication-incompetent adenovirus vector, could selectively induce growth suppression and apoptosis in HCC cell line SMMC-7721 but without any toxic side-effect on normal liver line L02.

In Vivo Target RNA Specificity of Trans-Splicing Phenomena by the Group I Intron

The Tetrahymena group I intron has been shown to employ a trans-splicing reaction and has been modified to specifically target and replace human telomerase reverse transcriptase (hTERT) RNA with a suicide gene transcript, resulting in the induction of selective cytotoxicity in cancer cells that express the target RNA, in animal models as well as in cell cultures. In this study, we evaluated the target RNA specificity of trans -splicing phenomena by the group I intron in mice that were intraperitoneally inoculated with hTERT-expressing human cancer cells to validate the anti-cancer therapeutic applicability of the group I intron. To this end, an adenoviral vector that encoded for the hTERT-targeting group I intron was constructed and systemically injected into the animal. 5'-end RACE-PCR and sequencing analyses of the trans-spliced cDNA clones revealed that all of the analyzed products in the tumor tissue of the virus-infected mice resulted from reactions that were generated only with the targeted hTERT RNA. This study implies the in vivo target specificity of the trans - splicing group I intron and hence suggests that RNA replacement via a trans -splicing reaction by the group I intron is a potent anti-cancer genetic approach.

Expression and Significance of Differential Expression Gene Hepsin in the Prostate Cancer / 中国康复理论与实践

@#Objective To evaluate the expression and significance of differential expression gene Hepsin in the prostate cancer (PC) screened by the cDNA microarray technique.Methods The techniques of semiquantitative RT-PCR and Western blotting were used to detect the mRNA and protein expression of Hepsin. Specimens of 40 cases of PC, 15 benign prostatic hyperplasia (BPH) and 6 normal prostate tissues were examined by the immunohistochemical stain.Results Hepsin was more expressed in PC tissue than normal prostate tissue (P=0.026) and was confirmed by Western blot analysis. Immunohistochemical test confirmed this and demonstrated that Hepsin did not expressed in normal prostate but expressed in PC and BPH and there was a significant difference in Hepsin expression level between PC and BPH tissues ( P=0.000).Conclusion Hepsin high expressed in PC may be a new molecular marker in early diagnosis of PC and a new target for gene therapy of PC.

Changing the Composition of Fatty Acids of Cell Membrane Promotes Breast Cancer Cell to Apoptosis / 中国生物工程杂志

Objective: To study expression of the gene of n-6 fatty acid desaturase fat-1 in human breast cancer cell, composition change of fatty acids of cell membrane, and effect of the gene on apoptosis of breast cancer cell. Methods: Construct recombinant adenovirus vector (Ad.GFP.fat-1) containing fat-1 gene, the recombinant adenovirus was produced in 293 package cell, then it infected the breast cancer cells MCF-7; Total RNA of the cells was isolated and hybridized with antisense RNA probe of fat-1 mRNA by Northern to analyze the expression of fat-1 in MCF-7;The effect of fat-1on the proliferation of MCF-7 cell was analyzed by MTT method,apoptosis of the cells were detected by apoptosis kit;Content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography. Results: The proposed recombinant virus was got through DNA recombinant technique; fat-1 gene effectively expressed in human breast cancer cell MCF-7; The fat-1 mRNA band appeared 2 days after infection of virus Ad.GFP.fat-1;Compared with the control cell (Ad.GFP), proliferation of MCF-7 cell was markedly inhibited by the gene fat-1( 23%, p

Enhanced Oncolytic Effect and Anti-Tumor Effect of Replication Competent Adenovirus with Double Mutation in E1A & E1B Regions

Gene-modified replication-competent adenoviruses (Ads) are emerging as a promising new modality for the treatment of cancer. We have previously shown that E1B 19kDa and E1B 55kDa gene deleted Ad (Ad-deltaE1B19/55) exhibits improved tumor-specific replication and cell lysis, leading to potent anti-tumor effect. As an additional effort to increase cancer cell-selectivity of replicating adenovirus, we have first generated eleven E1A-mutant Ads (Ad-mt#1~#11) with deletion or substitution in retinoblastoma (Rb) binding sites of E1A. Of these viruses, Ad-mt#7 demonstrated significantly improved cytopathic effect (CPE) and viral replication in a cancer cell-specific manner. To further increase the cancer cell-specific killing effect of Ad-mt#7, both E1B 19kDa and E1B 55kDa genes were deleted, resulting in an Ad-deltaE1Bmt7. As assessed using CPE assay, MTT assay, and immunoblot analysis for Ad fiber expression, Ad-deltaE1Bmt7 exerted markedly enhanced cancer cell-specific killing effect as well as viral replication in comparison to either Ad-mt#7 or Ad-deltaE1B19/55. Furthermore, the growth of established human cervical carcinoma in nude mice was significantly suppressed by intratumoral injection of Ad-deltaE1Bmt7. In summary, we have developed an oncolytic adenovirus with significantly improved therapeutic profiles for cancer treatment.

Development of Hepatoma-Specific Replication Competent Adenovirus

A prerequisite for the development of a cancer cell selective targeting adenovirus is the generation of adenoviral vectors that lack native receptor binding ability and additionally contain domains redirecting the vector to cancer cell specific receptors. Towards this goal, we have generated an E1B 55kDa-deleted oncolytic and coxoackie and adenovirus receptor (CAR)-binding ablated adenovirus, YKL-K420A. This newly engineered adenovirus resulted in a dramatic reduction of transduction efficiency compared to the control adenovirus, YKL-1, in all of the cell lines tested. The malaria circumsporozoite (CS) protein interacts with glycosaminoglycans (GAG) present on the liver cell surface, and plays a prominent role in sporozoite attachment and invasion into hepatocytes. To redirect the CAR binding ablated adenovirus YKL-K420A to hepatocytes, CS protein epitope (EWSPCSVTCGNGIQVRIK) was incorporated onto the C-terminus of the YKL-K420A fiber protein, generating an YKL-K420A-hepa. The In vitro efficacy and specificity of YKL-K420A-hepa was then evaluated by comparing the cytopathic effect in hepatoma and other cancer cells from different origins. In hepatoma cells, YKL-K420A-hepa exerted upto 20-fold higher cytolytic ability compared to the control adenovirus, YKL-1, in hepatoma cell lines. Treatment with YKL-K420A-hepa also significantly suppressed tumor growth in a hepatoma xenograft tumor model when compared to YKL-1. Taken together, these studies demonstrate that the strategy to alter adenovirus tropism may greatly improve adenoviral utilities in gene therapy applications.

Application and Prospection of RNAi in Colorectal Cancer Gene Therapy / 中国普外基础与临床杂志

Objective To explore the application of RNA interference(RNAi) in colorectal cancer gene therapy.Methods The related literatures in recent years were reviewed.Results RNAi causes a high effective and distinctive degradation of mRNA homologous in sequence to the dsRNA.This new technology has been successfully applied to research the genesis and the growth of colorectal cancer.Conclusion RNAi has been a new focus in gene therapy for colorectal cancer.