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Hepatocellular carcinoma is one of the most common malignancies and leading cancer-associated deaths worldwide. Ozone has been proposed as a promising therapeutic agent in the treatment of various disorders. The purpose of this paper is to assess the potential anticancer effects of the ozone on liver cancer cells. The liver cancer cell line of bel7402 and SMMC7721 was used in this study. Proliferation was evaluated using the CCK-8 and the colony formation assay. Wond healing assay and transwell assay without Matrigel were used to evaluate their migration ability. Flow cytometry was used for cell cycle analysis and reactive oxygen species (ROS) determination. Glutathione detection kit was used for measurement of glutathione level. Protein expression was estimated by western blot analysis. Ozone treatment inhibited liver cancer cell proliferation, colony formation. Ozone induced G2/M phase cell cycle arrest, which could be elucidated by the change of protein levels of p53, p21, Cyclin D1, cyclin B1, cdc2, and CDK4. We also found that ozone treatment inhibited migration ability by inhibiting EMT-relating protein. Ozone also induced ROS accumulation and decreased glutathione level decreased, which contributed to the inactivation of the PI3K/AKT/NF-κB pathway. Finally, we found that pre-treatment of liver cancer cells with N-acetylcysteine resisted ozone-induced effects. Ozone restrains the proliferation and migration potential and EMT process of liver cancer cells via ROS accumulation and PI3K/AKT/NF-κB suppression.

Background. The treatment of cervical cancer in the late stage is still quite challenging, because of nonspecificity in conventional therapies and the lack of molecular targeted drugs. It is necessary to find novel biomarkers for cervical cancer treatment. Methods. In the present study, cervical cell lines HeLa and SiHa with kin17 knockdown were constructed by transfection of the recombinant lentiviral vector carrying KIN17 siRNA and screened by puromycin. The established cells with kin17 knockdown were determined by fluorescence observation and western blotting. Cell apoptosis and the mitochondrial membrane potential (MMP) were detected by flow cytometry. The activity of caspase 3 enzyme was tested by spectrophotometry. The expression profile of apoptosis-associated proteins was analyzed by western blotting. Finally, we used bioinformatics and proteomic data to analyze KIN-related genes in cervical cancer. Results. The results showed high fluorescent positive rates (>90%) and high gene silencing efficiency (>65%) in HeLa and SiHa cells transfected with gene silencing vectors. Moreover, kin17 deficiency decreased the MMP and increased the apoptosis rates in HeLa and SiHa cells, respectively. Furthermore, knockdown of kin17 enhanced the activity of caspase 3 enzyme, increased the expression of cleaved PARP and Bim, while decreasing the expression of Bcl-xL and phosphorylated BAD in HeLa and SiHa cells. Identification of KIN-related prognostic genes in cervical cancer revealed that a total of 5 genes (FZR1, IMPDH1, GPKOW, XPA, and DDX39A) were constructed for this risk score, and the results showed that CTLA4 expressions were negatively correlated with the risk score. Conclusion. Our findings demonstrated that kin17 knockdown facilitates apoptosis of cervical cancer cells by targeting caspase 3, PARP, and Bcl-2 family proteins. Besides, kin17 could regulate cancer cell apoptosis through the mitochondrial pathway and could be used as a novel therapeutic target for the regulation of cell apoptosis in cervical cancer.

Guangdong is a main dengue epidemic area in China and undergoes dengue outbreaks frequently. In 2014, Guangdong experienced a serious dengue outbreak with 45,224 confirmed cases and six deaths, which might affect the seroprevalence among the local population. There is evidence that dengue virus (DENV) infection during pregnancy may cause adverse outcomes. Therefore, it is important to assess the seroprevalence of DENV among Guangdong pregnant women. We aimed to survey the seroprevalence of anti-DENV antibodies among pregnant women in Guangdong and to analyze the features of different seroprofiles. We collected a total 951 samples from pregnant women living in Guangdong in 2016. All serum samples were screened for DENV-specific antibodies (IgG and IgM) by indirect enzyme-linked immunosorbent assay (ELISA) and confirmed using capture ELISA. In IgM-positive samples, we performed DENV RNA detection using reverse transcriptase polymerase chain reaction. We collected information of delivery outcomes, neonate features, and clinical laboratory variables of parturients and newborns following delivery. Seroprevalence of DENV among the women in our sample was 4.31%. A total 1.26% and 3.15% of samples were IgM and IgG positive, respectively. In addition, 22.22% of IgM-positive and 9.09% of IgG-positive participants had adverse outcomes. There was no difference with respect to adverse outcomes compared with controls (8.80%) who were IgG and IgM negative. There was no difference in clinical laboratory variables among the different seroprofiles. We found a high seroprevalence of DENV among pregnant women in Guangdong comparing with the overall seroprevalence of local population before 2014. Asymptomatic DENV infection during pregnancy was not found to contribute to adverse outcomes.

BACKGROUND: Oxidative stress is an important factor in the modulation of both tumorigenesis and anticancer responses. Ozone (O(3)) is a strong oxidant that causes redox reactions and exerts anticancer effects in various types of cancer cells. However, the pathways involved in O(3)-induced cell death are not well understood. METHODS: In vitro human hepatocellular carcinoma (HCC) BEL7402 cells were treated with various O(3) concentrations to evaluate O(3) cytotoxicity by Cell Counting Kit-8 (CCK-8) assay and flow cytometry. The regulatory mechanisms were analyzed by western blot analysis. In vivo, an HCC model was established to evaluate the inhibition of HCC with O(3) treatment. RESULTS: In vitro cells treated with O(3) exhibited a round and small morphology with nuclear shrinkage and fragmentation. The CCK-8 assay confirmed the potent cytotoxic activity of O(3) against BEL7402 cells (IC(50) value of 5 µg/mL). Acridine orange/ethidium bromide (AO/EB) staining revealed apoptosis of BEL7402 cells after O(3) treatment. Flow cytometry analysis showed that S phase cell cycle arrest and apoptosis increased with O(3) exposure. In addition, O3 exposure reduced the mitochondrial membrane potential (ΔΨm) and induced reactive oxygen species (ROS) accumulation. Western blot analysis showed that O(3) exposure reduced B-cell lymphoma 2 (BCL-2) expression and increased cleaved poly ADP-ribose polymerase (PARP), cytochrome C (Cyt-C), caspase-3, caspase-9, and p-JNK expression. In vivo, treatment with intratumor injection O(3) (20 µg/mL) inhibited HCC growth. CONCLUSIONS: Overall, our findings showed that O(3) induces BEL7402 cell apoptosis via the intrinsic mitochondria-dependent pathway. Therefore, O(3) has therapeutic potential for HCC.

Effective therapy for breast cancer has been extensively studied worldwide, particularly for triple-negative breast cancer and drug-resistant subtypes. DNA/RNA-binding protein KIN17 (kin17) has been reported to be significantly upregulated in breast cancer cells, and is proposed to serve a role in the regulation of cell proliferation. The present study further investigated the association of kin17-knockdown with breast cancer cell apoptosis. Cell Counting kit-8, flow cytometry, TUNEL assay and caspase 3/7 analysis were performed on MDA-MB-231 cells to determine the association between kin17 and breast cancer cell apoptosis. In addition, western blot analysis was performed to investigate the mechanism of kin17 in the apoptosis of MDA-MB-231 cells. The results revealed that knockdown of kin17 inhibited proliferation and promoted apoptosis of MDA-MB-231 cells, and suggested a poly (adenosine diphosphate-ribose) polymerase-related mechanism behind the apoptosis of the cells. These findings suggested that kin17 could become a novel target for breast cancer therapy.