Hepatitis B virus X protein-mediated non-coding RNA aberrations in the development of human hepatocellular carcinoma

Hepatitis B virus (HBV) has an important role in the development of human hepatocellular carcinoma (HCC). Accumulated evidence has shown that HBV-encoded X protein (HBx) can induce both genetic alterations in tumor suppressor genes and oncogenes, as well as epigenetic aberrations in HCC pathogens. Non-coding RNAs (ncRNAs) mainly include microRNAs and long non-coding RNAs (lncRNAs). Although ncRNAs cannot code proteins, growing evidence has shown that they have various important biological functions in cell proliferation, cell cycle control, anti-apoptosis, epithelial–mesenchymal transition, tumor invasion and metastasis. This review summarizes the current knowledge regarding the mechanisms and emerging roles of ncRNAs in the pathogenesis of HBV-related HCC. Accumulated data have shown that ncRNAs regulated by HBx have a crucial role in HBV-associated hepatocarcinogenesis. The findings of these studies will contribute to more clinical applications of HBV-related ncRNAs as potential diagnostic markers or as molecular therapeutic targets to prevent and treat HBV-related HCC.

Expression of HDAC4 in human lung adenocarcinoma and its relationship with chemotherapy resistance / 医学研究生学报

Objective There have been few research on the relationship between expression of HDAC4 and chemotherapy resistance in human lung adenocarcinoma.The present study aims to investigate the expression and clinical significance of HDAC4 in human lung adenocarcinoma tissues.Methods We selected 72 tissues in lung adenocarcinoma patients with docetaxel-resistant from January 2006 to December 2007 in Department of Oncology and Thoracic Surgery, Nanjing General Hospital of Nanjing Military Region, then evaluated the recent efficacy according to the RECIST criteria and divided the tissues into sensitive(n=32, included complete remission and partial remission) and insensitive(n=40, included stability and progress) groups.The expression of HDAC4 in tissues≥the HDAC4 optimal relative expression cut-off value(78.7) was high level HDAC4 group(n=35), otherwise it was low level HDAC4 group(n=37).QRT-PCR analysis was performed to detect the HDAC4 expression levels in sensitive group and insensitive group.Analyzed the progression free survival in high level HDAC4 group and low level HDAC4 group.Results The expression of HDAC4 was significantly higher in the insensitive group compared with the sensitive group [(1.42±0.30) vs (0.60±0.15), P<0.01].The median progression free survival was significantly shortened in the high level HDAC4 group compared with the low level HDAC4 group (10.2 months vs 5.8 months, P<0.05).ConclusionThe expression of HDAC4 increased in docetaxel-resistant lung adenocarcinoma patients, and it is expected to be a predictive indicator of the resistance of docetaxel.

Aurora-A/NF-κB is involved in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells / 医学研究生学报

Objective Human lung adenocarcinoma SPC-A1/DTX cells have a higher radioresistance than SPC -A1cells. This study was to investigate the role of Aurora-An/uclear factor κB ( NF-κB) in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells and its possible molecular mechanisms . Metho ds We collected human lung adenocarcinoma SPC-A1 and SPC A1/DTX cells and divided them into four groups:sh-Aurora-A ( Aurora-A plasmid interference ) , sh-NC, NF-κB inhibition ( SPC-A1/DTX +NF-κB inhibitor ) , and DMSO control .We measured the in vitro radio-sensitivity of the cells by MTT assay , determined their proliferation ability by cloning assay , and detected the mRNA and protein expressions of the target genes by real -time quantitative RT-PCR and Western blot , respectively . Results The 50% effective doses ( ED50 ) of the SPC-A1 and SPC-A1/DTX cells on radiotherapy were (6.5 ±0.3) and (12.8 ±0.6) Gy, respectively, with statisti-cally significant difference between the two groups ( P <0.01 ) .In the radiation doses of 0, 2, 4, and 6 Gy, the numbers of the cloned SPC-A1 cells were 345 ±20 , 252 ±22 , 170 ±15 , and 81 ±10 , sig-nificantly lower than those of the cloned SPC -A1/DTX cells (402 ±21, 370 ±18, 301 ±16, and 252 ±15) (P<0.05).The protein and mRNA expressions of Aurora-A were remarkably higher in the SPC-A1/DTX than in the SPC-A1 cells (1.00 ±0.08 and 1.00 ±0. 06 vs 0.49 ±0.03 and 0.22 ±0.02, P<0.05).MTT assay showed a higher ED50 in the sh-NC than in the sh-Aurora-A cells ([11. 8 ±0.5] vs [7.1 ±0.3] Gy, P<0.01) as well as in the control than in the NF-κB inhibition group ([11.7 ±0.5] vs [6.1 ±0.3] Gy, P<0.01).Inhibition of Aurora-A increased the expression of IκBa by 2.18 ±0.32 times (P<0.01) and that of NF-κB by 0.24 ±0.03 times (P<0.01).The expressions of IκBa (1.00 ±0.05) and NF-κB (1.00 ±0.04) were significantly lower in the parent strains of SPC-A1 than 0.65 ±0.04 and 2.18 ±0.15 in the drug-resistant strains of SPC-A1/DTX (P<0.01). Conclusi on Auro-ra-A/NF-κB is involved in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells.

Genome-wide screening of CpG island methylation changes induced by low-dose X-ray radiation in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the methylation changes in promoter CpG islands induced by low-dose X-ray radiation (LDR).</p><p><b>METHODS</b>Twenty male BALB/c mice were randomly divided into control and fractionated radiation group exposed to 6 MV X-ray for 10 days (0.05 Gy/day). All the mice were sacrificed 2 h after the last radiation on day 10, and blood samples were collected for detecting DNA methylation changes using Roche-NimbleGen mouse DNA methylation 3×720K Promoter Plus CpG Island Array. MeDIP-qPCR was used to further validate the methylation status of specific genes.</p><p><b>RESULTS</b>A total of 811 genes were found to show specific hypermethylation in fractional radiation group as compared with the control group, involving almost all the main biological processes by GO analysis. Eight candidate genes (Rad23b, Tdg, Ccnd1, Ddit3, Llgl1, Rasl11a, Tbx2, and Slc6a15) were confirmed to be hypermethylated in LDR samples by MeDIP-qPCR, consistent with the results of the methylation chip study.</p><p><b>CONCLUSION</b>LDR induces promoter hypermethylation on specific genes, which may contribute to radiation-induced pathogenesis.</p>

The fork head box M1 effects on human colon cancer cells malignant phenotype / 医学研究生学报

Objective The invasion and metastasis of colon cancer often leads to treatment failure and mortality in patients . Our research is to investigate the influence of FoxM 1 to malignant human colon cancer line . Methods In two human colon cancer lines, the protein and mRNA expression levels of FoxM 1 were analyzed with the application of RT-PCR and Western blot , from which high-expressed HT-29 and low-expressed HCT-116 were determined.The expression of FoxM1 was down-regulated by RNA interfering in HT-29 and up-regulated by constructing overexpression transgenic line in HCT-116.The proliferation of the above cells was assayed by healing method;while the metastasis and invasion ability were examined by Transwell chamber assay . Results Two colon cancer lines were selected with high-expression or low-expression of FoxM1 separately named HT-29 and HCT-116.Application of PEX-2-FoxM1 raised after HCT-116 cells express FoxM1, cell scratches in HCT-116 experimetal group ([70.92 ±1.48]%) compared with HCT-116 control group([16.92 ±4.05]%)and HCT-116 blank control group([16.66 ±2.63]%) will markedly enhance its capabil-ity of healing (P<0.05), Transwell Chambers in membrane cells in HCT-116 experimetal group (186.0 ±6.8) compared with HCT-116 control group(42.0 ±2.0) and HCT-116 blank control grou (37.0 ± 2.2)was increased (P<0.05).On the other hand, the applied pG-PH-shFoxM1 can reduce FoxM1 expression in HT-29 cell, cell scrat-ches healing ability in HT-29 experimetal group ( [ 10 .37 ± 3.86]%) compared with HT-29 control group([34.63 ±2.35]%)and HT-29 blank control group([67.36 ±2.61]%) decreased significantly (P<0.05), Transwell Chambers in membrane cells in HT-29 experimetal group (53.0 ±1.8)compared with HT-29 control group(95.0 ±2.2)and HT-29 blank control grou(118.0 ±4.0) was also reduced (P<0.05). Conclusion The expression of FoxM1 is in close relation to the invasion and metastasis of CRC .The fact that the siRNA interfering FoxM1 could effectively inhibit the proliferation, metastasis and invasion, suggesting FoxM1 could po-tentially be a new molecular target for inhibiting the proliferation of human colon cancer line .

Study on diallyi disulfide inhibiting the proliferation of human small cell lung cancer H446 cells / 中国医师杂志

Objective To investigate the effects of diallyl disulfide (DADS) on ceil proliferation in human small cell lung cancer H446 cells in vitro. Methods MTT assay was used to observe inhibitory effect of DADS on proliferation of H446 cells. Cell Proliferation in-hibition was measured by growth curve analysis, average doubling time, vitality detection and MT]" assay. Cell morphology was observed by inversion microscope and optics microscope. Cell apoptosis was analyzed by cell morphology observed under light microscope, flow cytometry (FCM). Results MTT assay showed that DADS from 4 to 60 μg/ml significantly inhibited t446 ceils and exhibited a dose-dependent and time-dependent model. After exposure to DADS, H446 cell average doubling time retarded from 25. 40 hours to 145. 64 hours( P＜0.05).Flow cytometry analysis revealed that the cell content of C0-phase declined, however, hypodiplod peak was increased, which means cell ap-optosis were induced by DADS. Conclusion DADS could significantly inhibit the proliferation of H446 cells and induce the apoptosis of H446 cell.

Screening differential expression of docetaxel-resistance related genes of human lung adenocarcinoma cell line SPC-A1 by cDNA microarray / 中国肺癌杂志

<p><b>BACKGROUND</b>Docetaxel is one of effective chemotherapeutics in the last few years, however, it is interfered by drug resistance in its further application. The aim of this study is to screen differentially expressed genes of docetaxel resistant cell line SPC-A1/Docetaxel and its parent cell line SPC-A1 with gene chip technique.</p><p><b>METHODS</b>The cDNA retro-transcribed from equal quantity mRNA derived from SPC-A1/Docetaxel and SPC-A1 cell lines. The mixed probes were hybridized with Affymetrix GeneChip HG-U133A2.0. The acquired image was analyzed by Affymetrix GeneChip Operating Software Version 1.0. Then, part of these results were verified by RT-PCR.</p><p><b>RESULTS</b>A total of 934 differentially expressed genes were screened out, in which up-and down-regulated genes were 428 and 506 respectively. These genes involved in ABC transporter, apoptosis regulator, tubulin, signal transducer, enzyme and so on.</p><p><b>CONCLUSIONS</b>These differentially expressed genes may be related to the mechanisms of docetaxel resistance in SPC-A1/Docetaxel cell line.</p>

The Cross-talk Between p27~(kip1) and Its Interacting Molecules in Breast Cancer Cells / 生物化学与生物物理进展

p27kip1 is an important negatively regulator of cell cycle progression and plays a central role in the pathogenesis of a member of tumors including breast cancer. In breast cancer cells, the level of p27kip1 expression usually decreases during tumor development and progression, in addition, cytoplasm mislocalization of p27kip1 has been reported, but less is known about the exact molecular mechanisms. Studies have indicated that phosphorylation is the key regulation way, several signal transduction pathways are involved in the regulation of the expression and distribution of p27kip1. To further understand the mechanism, the disparity of the interacting protein profiling between tumor cells and normal cells must be identified first. Including cyclins, cyclin-depend kinases, CRM1, jab1, SKP2, p27kip1 has various interacting molecules. There are also several interacting molecules especially for breast cancer cells. It seems that different protein profiling cause the different expression and intracellular distribution in different cell cycle phase. So, disparity of the p27kip1 protein profiling may be the main mechanism of its down-expression and mislocalization in breast cancer cells.

Expressions of survivin and caspase-3 in human non-small cell lung cancer and their relationship with cell apoptosis / 中国肺癌杂志

<p><b>BACKGROUND</b>Survivin, a member of inhibitor of apoptosis protein (IAP) family, can directly inhibit caspase-3 and caspase-7 activity and plays an important role in oncogenesis. The aim of this study is to investigate the expressions of survivin and caspase-3 in human non-small cell lung cancer (NSCLC), and to evaluate their relationship with cell apoptosis.</p><p><b>METHODS</b>The expressions of survivin and caspase-3 in 88 patients with NSCLC were examined by using immunohistochemical SP methods, and TNUEL method was used to detect the cell apoptosis simultaneously.</p><p><b>RESULTS</b>The positive expression rates of survivin in NSCLC tissues and normal lung tissues were 61.4% (54/88) and 13.8% (4/29) respectively, there was a significant difference between them (P < 0.01). Survivin expression in NSCLC was not related to the histologic type, pathological grade and lymph node metastasis (P > 0.05), but correlated with TNM stage (P < 0.05). The positive expression rate of caspase-3 was 89.7% (26/29) in normal lung tissues, which was higher than that in NSCLC tissues (73.9%, 65/88), but there was no significant difference (P > 0.05). Caspase-3 expression in NSCLC was associated with pathological grade (P < 0.05). The average apoptosis index (AI) of survivin-positive cases was significantly higher than that of surviving-negative ones (1.63±0.58 vs 3.29±0.76)(P < 0.05). The average AI of the caspase-3 positive cases was significantly higher than that of the caspase23 negative cases (2.42±0.59vs1.28±0.65)(P < 0.05). Expression of survivin was negatively correlated with caspase-3 (P < 0.05).</p><p><b>CONCLUSIONS</b>Survivin may play an important role in the process of carcinogenesis of NSCLC by inhibiting cell apoptosis. Moreover, survivin may prevent cell apoptosis by inhibiting caspase-3 activation.</p>

Immunomodulatory features of chemotherapeutics / 医学研究生学报

Chemotherapeutic drugs conventionally used for their cytotoxic activity can also exert their influence on the immune system by grooming the tumor microenvironment,abrogating immune tolerance and inciting immune reconstitution.Due to their immunomodulatory features,chemotherapeutics,used as a preconditioning regimen and combined with subsequent immunotherapy,can mobilize the immune system to generate antitumor immune response.The synergy between chemotherapy and immunotherapy has brought some enlightenment to the development of new protocols for cancer treatment.

Update of the diagnosis and treatment of intravascular large B-cell lymphoma / 医学研究生学报

Intravascular large B-cell lymphoma(IVLBCL) is a rare form of diffuse LBCL characterized by preferential intravascular growth of malignant lymphocytes,aggressive behavior,and an often fatal course.It displays some differences in clinical presentation among diverse geographical areas.The immunophenotype plays a critical role in diagnosis and differential diagnosis.Recent therapeutic approaches,such as the rituximab plus CHOP regimen,could have a positive impact in IVLBCL patients.The use of high-dose chemotherapy supported by autologous stem-cell transplantation may improve current outcomes.

Influence of radiosensitivity by mild hyperthermia in a pulmonary adenocarcinoma cell line / 医学研究生学报

Objective:To investigate the radiosensitization and the cell-cycle of mild hyperthermia(≤42℃)on human pulmonary adenocarcinoma cell line SPC-A-1 in vitro. Methods: The human pulmonary adenocarcinoma cell line SPC-A were treated with radiation and the combination of radiation with mild hyperthermia. Radiosensitivity was determined by clonogenic assay and quantified by calculating the thermal enhancement ratio (TER). Flow cytometry was used to observe the cell-cycle. Results: Do, Dq calculated from the dose-response curve for radiation combined with 41.5℃ were 1.390 Gy, 1.426 Gy, whereas 1.693 Gy, 2.453 Gy for radiation alone, respectively. TER was 1.218. The proportion of cells in S phase was found to be 14.81% in the radiation group. The values, after 48 hours and 72 hours, with 6Gy radiation combined immediate 41.5℃ one hour mild hyperthermia, were 5.89% and 9.08%, respectively, versus 18.8% and 31.91% with 6 Gy radiation alone. Conclusion:Radiosensitization of mild hyperthermia in SPC-A-1 cells associated with the hyper-radiosensitization of the cells in S phase.

Nonmyeloablative allogeneic hematopoietic stem-cell transplantation a new immunotherapy for solid tumor / 医学研究生学报

Based on the practive of traditional allogeneic hematopoietic stem-cell transplantation therapy allo-HSCT and the development of nonmyeloablative conditioning, a new immunotherapy for solid tumor nonmyeloablative allo-HSCT, has been initiated. The nonmyeloablative,preparative regimens are associated with the use of lower dose of drugs,and less treatment-related toxicities. Instead of achieving maximal tumor reduction, the regimens are disigned to induce adequate immunosuppression to permit the engraftment of donor hematopoietic stem cells and serve as the platform for the administration of donor T cell in adoptive cell therapy. Donor's T-cell mediates a graft-versus-tumor(GVT)effect. This response is effective for the eradication of acceptor's tumor cell. This article reviews the concept, rationale, early clinical results, and limitation of nonmyeloablative allo-HSCT as a novel immunotherapy in solid tumors.

Expressions of NOS2 and NOS3 in human non-small cell lung cancer and the relationship with tumor angiogenesis and lymph node metastasis / 医学研究生学报

Objective: To investigate the expression and distribution of NOS2 and NOS3 in human non-small cell lung cancer (NSCLC),and to evaluate the relationship between their expression and tumor angiogenesis and lymph node metastasis. Methods: the expression of NOS2, NOS3 and IMVD in 95 patients with NSCLC were examined using immunohistochemical methods (S-P), and the relationship between them and many clinicopathological parameters was analyzed. Results: The positive expression of NOS3 was associated with histological subtype, IMVD and lymph node metastases of NSCLC(P

Progress in the study of gene polymorphism and gastric carcinoma / 医学研究生学报

s: Gastric carcinoma is the most common malignant tumor in China. When exposed to the same susceptible factor, different people have different incidence. Recent studies revealed that it may be related to some genes' polymorphism. In this article, the polymorphism of metabolism enzyme gene, oncogene, anti-oncogene, immune factor gene, pepsinogen C gene and mucin gene are reviewed. We highlight the situation of gastric carcinoma with some susceptible genetypes, also the possibility of using them to help scanning.

Prospects for early diagnosis of prostate cancer by detection of plasma DNA / 中华男科学杂志

A major problem for the early diagnosis of prostate cancer is the lack of clinically useful tests for screening a preclinical and asymptomic population without resort to invasive diagnostic procedures. Recent studies have demonstrated the possibility to detect genetic alterations in plasma or serum DNA from patients with prostate cancer or other cancers. Quantification and molecular event are associated with advanced stages and circulating tumor cells. These results indicate a new approach to the early diagnosis and monitoring of prostate cancer by non-invasive screening procedures based on the analysis of genetic changes in plasma.

Angiogenesis and its influrencing factors in non-small cell lung cancer / 医学研究生学报

Objective: To study angiogenesis and its influencing factors in human non-small cell lung cancer(NSCLC). Methods: The values of intratumor microvessel density(IMVD) in 95 cases of NSCLC were determined and its correlation with pathological parameters was analyzed. The relationship between IMVD and expressions of MMP-2,MMP-9,TIMP-2,VEGF,b-FGF,CD44v6,NOS2,NOS3 and E-cad in NSCLC was assessed. Results: There was active tumor angiogenesis of NSCLC, and the values of IMVD were correlated with clinicopathological stages of NSCLC and lymph nodes metastasis. The values of IMVD in positive expressions of VEGF, b-FGF, MMP-2, TIMP-2,CD44v6,NOS2 and NOS3 were significantly higher than that in negative expressions, opposed to the expression of TIMP-2 and E-cad. Conclusion:IMVD is a valuable marker for evaluating stages and metastasis of NSCLC. MMP-2, TIMP-2, VEGF, b-FGF, CD44v6, NOS2, NOS3 and E-cad may take part in the regulation of angiogenesis in NSCLC.

Cancer therapeutic vaccines targeting hTERT-derived T-cell epitope peptide: Advances in researches / 医学研究生学报

Human telomerase reverse transcriptase(hTERT),a catalytic subunit of telomerase,is highly expressed in a vast majority of cancer cells,but not in most normal adult cells,which makes it a broadly applicable molecular target for cancer immunotherapy.T cell epitope-based vaccines make use of T-cell epitopes from tumor antigens,and have its own unique advantages in cancer immunotherapy.The T-cell epitope peptide vaccine derived from hTERT can efficiently evoke specific cytotoxic T lymphocytes(CTL) responses,and has exhibited its anti-tumor effect in vivo in mice.Now it has already been put into experiment on human beings and displayed a good application prospect.

The mechanism of the role of ABC transporters in the multi-drug resistance of SP cells in SPC-A1 cell lines / 医学研究生学报

Objective: Recent researches suggest that tumors are organized in a hierarchy of heterogeneous cell populations and that the capability of maintaining tumor formation/growth specifically resides in a small population of cells called cancer stem cells(CSCs).CSCs are resistant to traditional cancer chemotherapy and radiation therapy because they highly express ATP-binding cassette(ABC) drug transporters and are relatively quiescent,with higher DNA repair and anti-apoptosis abilities.The purpose of this study is to investigate the relationship of ABC transporters with the multi-drug resistance and cancer stem cells of SPC-A1 cell lines.Methods: On the basis of Docetaxel-resistant cell line-SPC-A1/ Docetaxel,we compared the content and biological characteristics of SP cells,the expression of ABC transporters and its effect on the multi-resistance to Docetaxel between SPC-A1 and SPC-A1/ Docetaxel cell lines.Results: SP cells existed in both the SPC-A1 and the SPC-A1/ Docetaxel cell line,with a higher content in the latter.P-gp and BCRP obviously expressed in the SPC-A1/ Docetaxel and SPC-A1/ Docetaxel-SP cells,but only the expression of BCRP was increased in SPC-A1-SP cells.SPC-A1-SP cells also showed obviously higher abilities of proliferation,cloning and tumor formation than SPC-A1/ Docetaxel-SP cells.The multi-drug resistance of SPC-A1/ Docetaxel cells could be reversed with verapamil,but their resistance to Docetaxel could not be reversed completely with the same concentration of the drug.Conclusion: BCRP plays a major role in separating SP cells with tumor stem cell traits and its expression counts for the multi-drug resistance of SPC-A1-SP cells.

The experimental study of 5-FU on inhibiting angiogenesis / 医学研究生学报

Objective:To investigate the antiangiogenesis ability of 5 FU. Methods:Human umbilical vein endothelial cells were cultured primarily in vitro and influence of 5 FU on HUVEC proliferation was evaluated by MTT method. Chick chorilallantoic membranes(CAM) model was used to check whether the neovascularization of CAM could be suppressed in vivo . Results:5 FU both inhibited proliferation of HUVEC in vitro and suppressed angiogenesis of CAM in vivo at none cytotoxic doses. Conclusion: 5 FU demonstrated antiangiogenesis ability without obvious cytotoxicity.