BACKGROUND:Urinary system's most prevalent malignant tumor is bladder cancer. The enzyme pyrroline-5-carboxylate reductase 1 (PYCR1) has pro-tumorigenic characteristics. In the present study, the upstream and downstream regulatory mechanisms of PYCR1 in bladder cancer were investigated.METHODS:The relationship between the expression of PYCR1 in bladder cancer and its prognosis was analyzed using a bioinformatics technique. Plasmid transfection and small interfering RNA were utilized to overexpress and silence genes, respectively. Utilizing MTT, colony formation, EdU, and transwell assays, the proliferation and invasiveness of bladder cancer cells were evaluated. Employing an RNA pull-down experiment and RNA immunoprecipitation, the relationship between RNAs was analyzed. Fluorescence in situ hybridization, immunohistochemistry, and western blotting were used to detect protein expression and localization. Flow cytometry was used to identify reactive species (ROS) expression in cells. Mitophagy was detected using immunofluorescence.RESULTS:PYCR1 was highly expressed in bladder cancer tissue and was related with a poor prognosis for the patient. By binding to PYCR1, the antisense RNA lncRNA-RP11-498C9.13 prevented the degradation of PYCR1 and promoted its production. Down-regulation of lncRNA-RP11-498C9.13 and PYCR1 inhibited the proliferation and invasiveness of bladder cancer cells and decreased tumorigenesis. In addition, it was found that the lncRNA-RP11-498C9.13/PYCR1 axis promoted ROS generation and induced mitophagy in bladder cancer cells.CONCLUSIONS:We demonstrated that lncRNA-RP11-498C9.13 promoted bladder cancer tumorigenesis by stabilizing the mRNA of PYCR1 and promoted ROS-induced mitophagy. The lncRNA-RP11-498C9.13/PYCR1/mitophagy axis was anticipated to be a significant therapeutic target for bladder cancer.

PDF file - 297K, Tumor incidence and latency in MMTV-HA-14-3-3zeta transgenic and wild type mice (S1); Mammary tumor histology and latency in female MMTV-HA-14-3-3zeta transgenic mice (S2); TMA Analysis (S3); Confusion matrix of SVM (S4).

Using the data of listed firms in China’s A-share heavy pollution industry between 2008 and 2020, based on organizational theory, this study examines the impact of prospector-type firms and defender-type firms on environmental information disclosure quality. Empirical evidence shows that prospector-type firms reduce environmental information disclosure quality, compared with defender-type firms. After a series of robustness tests, the conclusion is still valid. This paper tests the impact mechanism of business strategy on environmental information disclosure quality and finds that financing constraints play a mediating effect in the relationship between business strategy and environmental information disclosure quality. This paper enriches and expands the literature in the field of influencing factors of environmental information disclosure quality and economic consequences of business strategy. At the same time, the conclusion of this paper has important reference significance for regulators to formulate policies to improve environmental information disclosure quality according to the heterogeneity of business strategy.

RNA polymerase mitochondrial (POLRMT) expression and the potential biological functions in skin squamous cell carcinoma (SCC) were explored. We showed that POLRMT is significantly elevated in skin SCC. Genetic depletion of POLRMT, using shRNA-induced knockdown or CRISPR/Cas9-mediated knockout (KO), resulted in profound anti-skin SCC cell activity. In patient-derived primary skin SCC cells or immortalized lines (A431 and SCC-9), POLRMT shRNA or KO potently suppressed mitochondrial DNA (mtDNA) transcription and suppressed cell viability, proliferation and migration. POLRMT shRNA or KO impaired mitochondrial functions in different skin SCC cells, leading to production of ROS (reactive oxygen species), depolarization of mitochondria and depletion of ATP. Moreover, mitochondrial apoptosis cascade was induced in POLRMT-depleted skin SCC cells. IMT1, a POLRMT inhibitor, largely inhibited proliferation and migration, while inducing depolarization of mitochondria and apoptosis in primary skin SCC cells. Contrarily, ectopic overexpression of POLRMT increased mtDNA transcription and augmented skin SCC cell growth. Importantly, POLRMT shRNA adeno-associated virus injection robustly hindered growth of the subcutaneous A431 xenografts in mice. In the POLRMT shRNA virus-treated A431 xenograft tissues, POLRMT depletion, mtDNA transcription inhibition, cell apoptosis, lipid peroxidation and ATP depletion were detected. Together, overexpressed POLRMT increases mtDNA transcription and promotes skin SCC growth.

Molecularly-targeted agents are still urgently needed for better non-small cell lung cancer (NSCLC) therapy. CC-115 is a potent DNA-dependent protein kinase (DNA-PK) and mammalian target of rapamycin (mTOR) dual blocker. We evaluated its activity in different human NSCLC cells. In various primary human NSCLC cells and A549 cells, CC-115 potently inhibited viability, cell proliferation, cell cycle progression, and hindered cell migration/invasion. Apoptosis was provoked in CC-115-stimulated NSCLC cells. The dual inhibitor, however, was unable to induce significant cytotoxic and pro-apoptotic activity in the lung epithelial cells. In primary NSCLC cells, CC-115 blocked activation of mTORC1/2 and DNA-PK. Yet, CC-115-induced primary NSCLC cell death was more potent than combined inhibition of DNA-PK plus mTOR. Further studies found that CC-115 provoked robust oxidative injury in primary NSCLC cells, which appeared independent of mTOR-DNA-PK dual blockage. In vivo studies showed that CC-115 oral administration in nude mice remarkably suppressed primary NSCLC cell xenograft growth. In CC-115-treated NSCLC xenograft tissues, mTOR-DNA-PK dual inhibition and oxidative injury were detected. Together, CC-115 potently inhibits NSCLC cell growth.

Early diagnosis of cervical cancer is essential for a good prognosis. Terahertz wave detection technology is a nondestructive and label-free physical detection technology, which can detect and monitor the cancer cells in real time, especially for patients with deep or inaccessible tumors. In this study, a single-cell-layer microfluidic device was developed. After replacing the optical clearing agent, the characteristics of H8, HeLa and SiHa cell lines in adherent and suspended states were detected. Additionally, the absorption increased with increasing cell density. For the mixed suspension cell samples, principal component analysis-support vector machine method was used to identify benign and malignant cell component. After living cells formaldehyde, changes in cell membrane permeability were evaluated to identify the cell survival status (i.e., dead or living) based on terahertz spectroscopy amplitude differences. Therefore, extending the terahertz spectrum detection to the molecular level can characterize the life essence of cells and tissues.

The PI3K/AKT signaling pathway plays an important role in tumor occurrence. PTEN can negatively regulate the PTEN/AKT signaling pathway, and DJ-1 is the negative regulator of PTEN. DJ-1 upregulation is associated with various tumor occurrences. Curcumin (Cur) is a phenolic compound extracted from various plant rhizomes with an antitumor pharmacological effect. This study investigated the role of Cur in regulating the DJ-1-PTEN/PI3K/AKT pathway activity and lung cancer cell proliferation and apoptosis. Human normal lung epithelial BEAS-2B cells and lung cancer A549 cells were cultured in vitro. Real-time quantitative PCR (qRT-PCR) was used to measure DJ-1 and PTEN mRNA expressions. Western blot was adopted to test DJ-1 and PTEN protein levels. A549 cells were treated with 0, 2.5, 5, and 10 μM of Cur. Cell proliferation activity was assessed using a CCK-8 assay and EdU staining. Cell apoptosis was determined by flow cytometry. A549 cells were divided into a control group, a 10 μM Cur treatment group, and a Cur+PTEN inhibitor SF1670 group. Compared with the BEAS-2B cells, DJ-1 was significantly increased, but the levels of PTEN were clearly decreased in the A549 cells. Cur treatment markedly inhibited A549 cell proliferation and induced apoptosis in a dose dependent manner. Cur downregulated DJ-1 levels and enhanced PTEN expression in A549 cells in a dose dependent manner. The combined administration of SF1670 enhanced the phosphorylation activity of p-AKT, attenuated the inhibitory effect of Cur on the p-AKT protein, and attenuated the pro-apoptotic and proliferative functions of Cur. Cur can inhibit lung cancer cell proliferation and promote apoptosis by downregulating DJ-1 to regulate the activity of PTEN/PI3K/AKT pathway.

MicroRNA-221 (miRNA-221) plays an important role in occurrence and development of tumor. However, expression pattern and role of miRNA-221 in the non-small-cell lung carcinoma (NSCLC) remains poorly understood. In the current study, we explored the roles and the underlying mechanism of miR-221 in NSCLC by gain and loss of function analysis. We found that the expression of miRNA-221 is significantly higher in NSCLC tissues and NSCLC cells compared to normal tissues and cells, respectively (P<0.05). NSCLC cell lines SPCA1 and H1299 were transfected with miRNA-221 mimic or inhibitor, respectively, and then performed to analysis. Experiments indicated that upregulation of miRNA-221 promotes cell proliferation, cell cycle, migration, invasion in SPCA1 cells, while downregulation of miRNA-221 exhibited the opposite role. Tissue inhibitor of metallopeptidases-2 (TIMP2) mRNA or protein levels were downregulated after miRNA-221 overexpression in SPCA1 and H1299 cells, respectively. Furthermore, TIMP2 was identified as a direct target of miRNA-221. In summary, our results indicate that miRNA-221 promotes NSCLC growth and invasion through repressing the expression of TIMP2, which suggest that inhibition of miRNA-221 could be a potential target for the treatment of NSCLC.

Radiotherapy is a major local treatment for cervical cancer.However, local uncontrollability due to radioresistance is still common.Therefore, the prediction of radiosensitivity is quite beneficial to develop an optimal treatment strategy for individual patients.Multiple factors could influence the radiosensitivity of cells, and p53 status is one of them.The upstream or downstream molecules of p53 could also be regulated to affect the radiosensitivity of cervical cancer.The aim of the review is to analyze the difference in p53 status between different types of cervical cancer and to discuss how p53 regulates the response to radiotherapy.

将地衣芽孢杆菌SR01(Bacillus licheniformis SR01)的植酸酶phy基因重组于表达载体pGEX-4T-3中,导入大肠杆菌BL21(Escherichia coli BL21)构建工程菌pGX1143,并得到了高效表达.对其酶学性质的研究表明:该酶最适反应pH为5.0,最适反应温度为55 ℃,具有广泛的pH稳定性和较好的热稳定性;Co2+、Li+、Mg2+和Ba2+对酶活性有促进作用,Cu2+、Pb2+、Fe2+、Ag+、SDS对酶活性有不同程度的抑制作用,并且Pb2+、Fe2+和SDS对酶活性的抑制作用较强烈;此外,该酶还具有非常好的抗胰蛋白酶水解能力和部分抗胃蛋白酶水解的能力.

OBJECTIVE:To investigate the reversal effect of targeted modulation of bcl-2 expression by miR-15a and miR-16 on drug resistance of human colon cancer cells.METHODS:Mimics or inhibitors of miR-15a and miR-16 were transfected into HCT8 or HCT8/VCR cells with the help of Lipofectamine 2000. The expressions of miR-15a and miR-16 mRNA were detected by RT-qPCR. The levels of bcl-2 and P-gp proteins were measured by Western blot. The inhibitory effects of VCR on growth of HCT8 and HCT8/VCR cells were detected by CCK8.RESULTS:After transfection of the mimics, the expression of miR-15a in the blank control group, negative control group and miR-15a mimic group was 1.00, 0.87 ± 0.24, and 223.44 ± 59.07, respectively, and miR-15a was increased significantly (P < 0.001). The expression of miR-16 in the blank control group, negative control group and miR-16 mimic group was 1.00, 0.66 ± 0.19, and 107.32 ± 22.58, respectively, and miR-16 expression was increased significantly (P < 0.001). The Western blot assay showed that the relative expressions of bcl-2 protein in the blank control group, negative control group, miR-15a mimic group and miR-16 mimic group were 1.00, 0.97 ± 0.02, 0.51 ± 0.06, and 0.65 ± 0.03, respectively, and the expression of bcl-2 protein was decreased significantly (P < 0.05), however, the expressions of P-gp protein showed no significant difference. The CCK8 test showed that at 1, 5, 25 and 125 µg/ml concentration of VCR, the survival rates of HCT8/VCR cells were basically the same in the blank control group, negative control group, miR-15a mimic group and miR-16 mimic group, but the survival rate of HCT8/VCR cells was significantly decreased after transfection of mimics (P < 0.05). After transfection of the inhibitors, the expressions of both miR-15a and miR-16 were decreased significantly (P < 0.001). The Western blot showed that the expression of bcl-2 protein was increased (P < 0.05), while the expression of P-gp protein showed no significant difference. The CCK8 test showed that the survival rate of HCT8 cells which were transfected with inhibitors was significantly higher than that of the blank control group (P < 0.05).CONCLUSIONS:miR-15a and miR-16 may reverse the drug resistance in human colon cancer cells. A possible mechanism is regulating the expression of bcl-2.