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.
Carcinoma, Hepatocellular* ; Cell Cycle Checkpoints ; Cell Proliferation ; Epigenomics ; Genes, Tumor Suppressor ; Hepatitis B virus* ; Hepatitis B* ; Hepatitis* ; Humans* ; MicroRNAs ; Neoplasm Metastasis ; Oncogenes ; RNA, Long Noncoding ; RNA, Untranslated*

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.

There is hypoxia in malignant glioma tissue,which would promote malignant progress and resistance to chemoradiotherapy.Hyperbaric oxygen can enhance dissolve oxygen in blood.Hyperbaric oxygen may restrain glioma growth through inhibition on cell proliferation,angiogenesis and promotion on apoptosis.Clinical research also found that combination of hyperbaric oxygen and chemoradiotherapy may ameliorate prognosis of glioma patient.But all results of previous studies still need further confirmation from multi-center,prospective,randomly controlled study.

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.

OBJECTIVETo study the methylation changes in promoter CpG islands induced by low-dose X-ray radiation (LDR).

METHODSTwenty 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.

RESULTSA 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.

CONCLUSIONLDR induces promoter hypermethylation on specific genes, which may contribute to radiation-induced pathogenesis.

Animals ; CpG Islands ; radiation effects ; DNA Methylation ; Dose-Response Relationship, Radiation ; Genome ; Male ; Mice ; Mice, Inbred BALB C ; X-Rays

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 .

Objective To study the methylation changes in promoter CpG islands induced by low-dose X-ray radiation (LDR). Methods 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. Results 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. Conclusion LDR induces promoter hypermethylation on specific genes, which may contribute to radiation-induced pathogenesis.

Objective To study the methylation changes in promoter CpG islands induced by low-dose X-ray radiation (LDR). Methods 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. Results 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. Conclusion LDR induces promoter hypermethylation on specific genes, which may contribute to radiation-induced pathogenesis.

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.

BACKGROUNDDocetaxel 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.

METHODSThe 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.

RESULTSA 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.

CONCLUSIONSThese differentially expressed genes may be related to the mechanisms of docetaxel resistance in SPC-A1/Docetaxel cell line.