MeDIP-seq reveals the features of mitochondrial genomic methylation in immature testis of Chinese mitten crab Eriocheir sinensis.

In this study, the methylation of mitochondrial genome in the immature testis of Chinese mitten crab Eriocheir sinensis of the Yangtze River system was determined for the first time using MeDIP-seq. Our methylated DNA fragments covered more than 99% of the mitochondrial genome in E. sinensis loaded from GenBank. There were 8 mutated bases and 42 SNPs in the crab mitochondrial genome. The methylation presented in all genes as well as in an A + T region, but less in intergenic regions in the mitochondrial genome. However, the level of methylation of most genes coding proteins and the A + T region were high. But, the majority of genes encoding tRNAs were hypomethylated, and both the rRNA genes also showed methylation of low or median frequency. Especially, the level of methylation of the intergenic regions is the lowest. Those features indicated that the methylation of DNA may play an important role in gene expressing regulation in the mitochondrial genome of immature testis in E. sinensis.

Molecular mechanisms of apoptosis in hepatocellular carcinoma cells induced by ethanol extracts of Solanum lyratum Thumb through the mitochondrial pathway.

AIM: To explore the induction effects and mechanism of Solanum lyratum Thumb (ST) on human hepatocellular carcinoma SMMC-7721 cells through the mitochondrial pathway. METHODS: The experiments were conducted on three groups: an experimental group (with ST ethanol extracts' concentration being 2.5, 5 and 10 mg/L), a negative control group (with only nutrient solution, 0 mg/L ST ethanol extracts), and a positive control group (2.5 mg/L DDP). The inhibition rate of cell proliferation was checked by using the methyl thiazolyl tetrazolium method, and cell apoptosis was tested by TUNEL method. Furthermore, RT-PCR was used to examine mRNA expression of Fas, FasL, caspase-8, caspase-3, p53 and Bcl-2 genes. RESULTS: Compared with the negative control group, the inhibition and apoptosis rates of the experimental group with different concentrations of ST extracts on human hepatocellular carcinoma SMMC-7721 cells significantly increased (P < 0.05). Besides, the mRNA expression of FasL and Bcl-2 significantly decreased (P < 0.05) while the mRNA expression of Fas, caspase-8, caspase-3 and p53 increased significantly. When compared with the positive control group, the experimental groups with 5 mg/L ST ethanol extracts showed effects similar to the positive control group. CONCLUSION: ST ethanol extracts induced the apoptosis of hepatocellular carcinoma SMMC-7721 cells through up-regulated Fas, caspase-8, caspse-3 and p53, and down-regulated FasL and Bcl-2 in the mitochondrial pathway.

Spautin-1 Ameliorates Acute Pancreatitis via Inhibiting Impaired Autophagy and Alleviating Calcium Overload.

Acute pancreatitis is characterized by zymogen pre-activation. Severe inflammation caused by zymogen activation can eventually lead to multiple organ dysfunctions, which contributes to the high mortality rate of severe acute pancreatitis. However, there is no specific treatment available for acute pancreatitis therapy. Here, we show that spautin-1, which effectively inhibits autophagy flux, ameliorated the pathogenesis of acute pancreatitis induced by cerulein or L-Arginine. CaMKII phosphorylation due to cytosolic calcium oeverload was revealed in this paper. It was also demonstrated that autophagic protein aggregates degradation blockade accompanying with impaired autophagy correlated positively to intra acinar cells digestive aymogen activation sitimulated by cerulein or L-Arginine. The role of spautin-1 in ameliorating acute pancreatitis was shown here to be associated with impaired autophagy inhibition and Ca2+ overload alleviation. We provided a promising therapy for acute pancreatitis here through targeting both impaired autophagy and increased cytosolic calcium.

[Solanum lyratum Thunberg alkaloid induces human lung adenoearcinoma A549 cells apoptosis by inhibiting NF-kappaB signaling pathway].

OBJECTIVE: To investigate the effects of solanum lyratum Thunberg alkaloid (STA) on induction of apoptosis and the expression of NF-kappaB signaling pathway related genes in A549 cells. METHODS: A549 cells was treated with STA in vitro. The proliferation inhibitory effect was evaluated by MTT assay. Induction of cell apoptotic rate was determined by flow cytometry method (FCM) after Annexin V-FITC/PI double staining. The expression of NF-kappaB/p65 in nuclei, Survivin, IkappaBalpha and p-1kaapaBalpha in cytosol were detected by western blot. RESULTS: STA exhibited strong proliferation inhibitory effect in a dose-and -time-dependent manner against A549 cells. After treated with STA for 24 h, the apoptotic rate was increased significantly. The expression of IkappaBalpha protein was increased markedly,while those of NF-kappaB/p65, Survivin and p-IkappaBalpha proteins were decreased markedly. CONCLUSION: STA can induce apoptosis of lung adenocarcinoma A549 cells, its mechanisms may be related to inhibition of NF-kappaB signaling pathway.

[Apoptosis-inducing effect of Scutellaria barbata extract on human lung cancer SPC-A-1 cells and the expression of apoptosis associated genes].

OBJECTIVE: To explore the apoptosis-inducing effect of Scutellaria barbata extract (SBE) and the expression of apoptosis associated genes survivin and caspase-3 on human lung cancer SPC-A-1 cells. METHODS: Lung cancer SPC-A-1 cells were treated with 2.5, 5 and 10 mg/L for 48 h,and the cells were treated with 2.5 mg/L DDP as positive control. The inhibitory rat was evaluated by MTT assay. Apoptotic rate was determined by TUNEL method. The expression of survivin and caspase-3 mRNA were detected by semi-quantitive RT-PCR. RESULTS: Compared with control group, the inhibitory rate was increased obviously (P < 0.001), the apoptotic rate was increased markedly (P < 0.01), the expression of caspase-3 mRNA was increased significantly (P < 0.05 or P < 0.01), while survivin mRNA was decreased markedly (P < 0.05 or P < 0.01) in SBE groups. CONCLUSION: SBE can induce apoptosis of SPCA-1 cells. The molecular mechanism may be related to up-regulating expression of caspase-3 and down-regulating expression of survivin genes.

[The influence of Solanum lyratum Thunb extract on apoptosis and the expression of fas/fasL genes in Hela cells].

OBJECTIVE: To explore the effects of Solanum lyratum Thunb (SL) extract on the apoptosis and the expression of fas and fasL genes in Hela cells. METHODS: The proliferation inhibitory rate was evaluated by MTF assay. Induction of cell apoptosis rate was determined by flow cytometry. The expression of Fas protein was detected by two-step immunhistochemical staining. The expression of fas and fasL mRNA was detected by semi-quantitive RT-PCR. RESULTS: SL extract displayed strong proliferation inhibitory effect in a dose-and-time-dependent manner against Hela cell. The rate of apoptosis was increased obviously. The expression of fas mRNA and protein was increased significantly, and fasL mRNA was decreased markedly. CONCLUSION: SL can induce apoptosis by up-regulating expression of fas and fasL genes, and inhibit the development of Hela cells.