N7-Methylguanosine Regulatory Genes Profoundly Affect the Prognosis, Progression, and Antitumor Immune Response of Hepatocellular Carcinoma.

Background: Hepatocellular carcinoma (HCC) is a common abdominal cancer with poor survival outcomes. Although there is growing evidence that N7-methylguanosine (m7G) is closely associated with tumor prognosis, development, and immune response, few studies focus on this topic. Methods: The novel m7G risk signature was constructed through the Lasso regression analysis. Its prognostic value was evaluated through a series of survival analyses and was tested in ICGC-LIRI, GSE14520, and GSE116174 cohorts. CIBERSORT, ssGSEA, and ESTIMATE methods were applied to explore the effects of the m7G risk score on tumor immune microenvironment (TIM). The GSEA method was used to evaluate the impacts of the m7G risk score on glycolysis, ferroptosis, and pyroptosis. The human protein atlas (HPA) database was used to clarify the histological expression levels of five m7G signature genes. The biofunctions of NCBP2 in hepatocellular cancer (HC) cells were confirmed through qPCR, CCK8, and transwell assays. Results: Five m7G regulatory genes comprised the novel risk signature. The m7G risk score was identified as an independent prognostic factor of HCC and could increase the decision-making benefit of traditional prognostic models. Besides, we established a nomogram containing the clinical stage and m7G risk score to predict the survival rates of HCC patients. The prognostic value of the m7G model was successfully validated in ICGC and GSE116174 cohorts. Moreover, high m7G risk led to a decreased infiltration level of CD8+ T cells, whereas it increased the infiltration levels of Tregs and macrophages. The glycolysis and pyroptosis processes were found to be enriched in the HCC patients with high m7G risk. Finally, overexpression of NCBP2 could promote the proliferation, migration, and invasion of HC cells. Conclusions: The m7G risk score was closely related to the prognosis, antitumor immune process, glycolysis, and malignant progression of HCC. NCBP2 has pro-oncogenic abilities, showing promise as a novel treatment target.

Prognostic and clinicopathological significance of PD-1 expression in hepatocellular carcinoma: a meta-analysis.

OBJECTIVE: We performed a meta-analysis to evaluate the prognostic and clinicopathological significance of programmed cell death-1 (PD-1) expression in patients with hepatocellular carcinoma (HCC). METHODS: We searched the Wanfang, Chinese Biomedical Literature, CNKI, PubMed, Embase, and Web of Science databases for relevant articles from inception to 1 July 2020. Statistical analysis was performed using RevMan 5.3 (Cochrane, London, UK) and Stata 14.0 software (StataCorp LP, College Station, TX, USA). RESULTS: Eight studies involving 732 patients with HCC were included. Higher expression of PD-1 predicted longer disease-free survival [hazard ratio (HR) 0.53, 95% confidence interval (CI): 0.38-0.72]. No significant correlation was observed between PD-1 expression and overall survival (HR 0.89, 95% CI: 0.58-1.35). PD-1 expression was correlated with age [odds ratio (OR) 0.66, 95% CI: 0.46-0.94] and alpha-fetoprotein level (OR 2.27, 95% CI: 1.45-3.55); no correlation was observed with sex, tumor size, tumor metastasis, hepatitis B virus history, tumor stage, or tumor multiplicity. Sensitivity analysis demonstrated no excessive effect on stability of the pooled results. No significant publication bias was found among the identified studies. CONCLUSION: PD-1 overexpression predicted better disease-free survival in patients with HCC. Moreover, PD-1 expression was associated with age and alpha-fetoprotein level.

A New Compound with Increased Antitumor Activity by Cotargeting MEK and Pim-1.

Feedback circuits are one of the major causes underlying tumor resistance. Thus, compounds that target one oncogenic pathway with simultaneously blocking its compensatory pathway will be of great value for cancer treatment. Here, we develop a new MEK inhibitor designated as KZ-02 that exhibits unexpectedly higher cytotoxicity than its starting compound AZD6244, a well-known MEK inhibitor, in colorectal cancer (CRC). Subsequent kinase selectivity study identified Pim-1 as an additional cellular target for KZ-02. Further studies showed that AZD6244 and Pim-1 1 (a Pim-1 inhibitor) have a synergistic effect on CRC suppression. Mechanistic study revealed that MEK inhibition by AZD6244 leads to increased Pim-1 expression, which could be a general mechanism behind the compromised cell-killing activity of MEK inhibitors. KZ-02, despite increasing Pim-1 mRNA expression, simultaneously promotes Pim-1 proteasomal degradation. Therefore, we uncover a new MEK inhibitor KZ-02 with significantly enhanced antitumor activity by co-targeting MEK and Pim-1.

Pretreatment with Lactobacillus fermentum XY18 Relieves Gastric Injury Induced by HCl/Ethanol in Mice via Antioxidant and Anti-Inflammatory Mechanisms.

AIM: Lactobacillus fermentum XY18 (LF-XY18) is a bacterial strain with satisfactory antioxidant properties in vitro that we previously isolated from Xinjiang yogurt. This article will explore the preventive effect of LF-XY18 on acute gastric injury and provide the basis for the innovative development and application of lactic acid bacteria (LAB). METHODS: Kunming mice underwent gastric injury induced by hydrochloric acid and ethanol. LF-XY18 isolated from yogurt in Xinyuan County in the Yili region of Xinjiang was subsequently administered intragastrically to mice for 2 weeks to explore the mechanism of LF-XY18 in preventing gastric injury via its antioxidant effects. RESULTS: There was decreased gastric juice volume, gastric injury area, and formation of gastric mucosal lesions in the LF-XY18 mice as compared to those in the control mice, while LF-XY18 prevented the decrease in the gastric juice pH value in mice. Compared with the gastric injury model group mice, LF-XY18 reduced the serum levels of motilin, substance P, interleukin-6, interleukin-12, tumor necrosis factor-α, and interferon-γ but increased the serum levels of somatostatin and vasoactive intestinal peptide. The activities of superoxide dismutase, glutathione peroxidase, glutathione, and nitric oxide were increased in the gastric tissue of the LF-XY18 mice compared with the control mice, but malondialdehyde activity was decreased in the LF-XY18 mice. Quantitative polymerase chain reaction analysis illustrated that in the gastric tissue of LF-XY18 mice, the messenger RNA (mRNA) expression of occludin, epidermal growth factor (EGF), EGF receptor, vascular EGF, inhibitor kappa-B-α, neuronal nitric oxide synthase, endothelial nitric oxide synthase, cuprozinc superoxide dismutase, manganese superoxide dismutase, and catalase was stronger than that in the control mice, but the mRNA expression of activated B cells (NF-κB), inducible nitric oxide synthase, and cyclooxygenase-2 was weaker than in the control mice. CONCLUSION: These results indicate that LF-XY18 has a potential role in the prevention of gastric injury through antioxidant effects, and a high concentration (1.0 × 109 CFU/kg b.w.) of LF-XY18 has a stronger anti-gastric injury effect than a low concentration (1.0 × 108 CFU/kg b.w.).

PAK: an essential motif for forming beta-turn structures and exhibiting the thrombolytic effect of P6A and its analogs.

Ala-Arg-Pro-Ala-Lys (ARPAK; also known as P6A) and 19 of its analogs were synthesized, and their thrombolytic activities were assessed in vitro and in vivo. The solution structures of 12 of the P6A analogs were determined using nuclear magnetic resonance (NMR) spectroscopy. The thrombolytic activity and conformational structure relationship was analyzed. We found that the Pro-Ala-Lys (PAK) sequence was essential for thrombolytic activity and was also responsible for the beta-turn structure found in the P6A analogs studied. The well defined beta turn may act as a binding head with the protruding lysine side-chain (positively charged) found at the target site for target recognition. Additionally, the N-terminal residue may be critical for thrombolytic activity, which for PAK-containing peptides, is likely achieved via a plasminogen-dependent pathway.

The composition and sequence specificity of Pro-Ala-Lys-OH for the thrombolytic activities of P6A and related oligopeptides.

The in vitro and in vivo thrombolytic activities of Ala-Arg-Pro-Ala-Lys-OH, its analogs and the related peptides were assayed. The results indicate that when (5)Lys of Ala-Arg-Pro-Ala-Lys-OH is changed into (5)Arg, and (3)Lys of Pro-Ala-Lys-OH is changed into (3)Arg the thrombolytic activities are collapsed; when Pro-Ala-Lys-OH is changed into Ala-Pro-Lys-OH, and Ala-Arg-Pro-Ala-Lys-OH is changed into Ala-Arg-Ala-Pro-Lys-OH the thrombolytic activities are also collapsed; when (5)Lys of Ala-Arg-Pro-Ala-Lys-OH is changed into (5)nLeu the thrombolytic activities are again collapsed. All of the results indicate that for the thrombolytic activities of Ala-Arg-Pro-Ala-Lys-OH and the related peptides Pro-Ala-Lys-OH exhibits either amino acid composition specificity or sequence specificity. The composition and sequence specificity of Pro-Ala-Lys-OH reflects its rule as the pharmacophore of P6A and the related peptides.

Synthesis and thrombolytic activity of carboline-3-carboxylic acid modified metabolites of Ala-Arg-Pro-Ala-Lys.

From the metabolism of H-Ala-Arg-Pro-Ala-Lys-OH, four metabolites, H-Pro-Ala-Lys-OH, H-Arg-Pro-Ala-Lys-OH, H-Ala-Arg-Pro-OH, and H-Ala-Arg-Pro-Ala-OH were identified. In order to find a new lead compound of thrombolytic peptide, 3S-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid was introduced to the N- and C-terminal of the metabolites by use of the common coupling strategy. Under this condition, the pseudopeptides (5a-d and 7a-d) were obtained with a good yield. The thrombolytic activities of 3S-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid containing oligopeptides were evaluated in vitro and in vivo. The result indicated that the thrombolytic activity of the pseudopeptide depended on the sequence and the modification pattern of the metabolites, and only when 3S-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid was introduced into the C-terminal of H-Pro-Ala-Lys-OH or H-Arg-Pro-Ala-Lys-OH, the desirable thrombolytic activity was retained and enhanced significantly.

[Role of matrix metalloproteinases (MMPs) in tumor invasion and metastasis: serial studies on MMPs and TIMPs].

We have conducted serial studies on the role of matrix metalloproteinases (MMPs), especially MMP-9, in tumor invasion and metastasis. In 9 human carcinoma cell lines derived from lung, prostate and melanoma, we found, by zymography and Western blot, that the expression levels of MMP-2 and MMP-9 correlated well with their invasive as well as metastatic abilities both in vitro and in nude mice. When anti-sense MMP-9 cDNA was introduced into WM451, a highly metastatic human melanoma cell line with high expression level of MMP-9, a significant down-regulation of MMP-9 protein expression was found. Meanwhile, the number of cells passing through Matrigel-coated membrane (in vitro invasion assay) and spontaneous metastases to lymph nodes and lungs were significantly reduced. Furthermore, when tissue inhibitors of metalloproteinases-1, -2 or -3 (TIMP-1, TIMP-2 or TIMP-3) cDNAs were individually transtected into metastatic cancer cells, remarkable inhibition of invasion and metastasis were also noticed in each group. These results demonstrate that either up-regulation of TIMPs or down-regulation of MMPs could significantly inhibit the expression of malignant phenotypes, suggesting the important role MMP-9 plays in tumor invasion and metastasis.

Effects of sex hormones and their balance on the proliferation of rat vascular endothelial cells.

OBJECTIVE: To investigate the adjustment of estrogen, progesterone and testosterone on the proliferation of female and male rat vascular endothelial cells (VECs) separately. METHODS: Rat lung VECs were cultured according to the block explanting method. MTT assay was used to measure the proliferation of VECs. RESULTS: 17beta-Estradiol (E(2)) at 3 x 10(-8) and 3 x 10(-7) M accelerated the proliferation of female rat VECs (p < 0.01). E(2) at 3 x 10(-9), 3 x 10(-8) and 3 x 10(-7) M accelerated the proliferation of male rat VECs (p < 0.05). Tamoxifen, the estrogen receptor antagonist, could block the effect of estrogen on the proliferation of VECs. Testosterone at 3 x 10(-8) and 3 x 10(-7) M significantly increased the proliferation of male rat VECs (p < 0.05), but had no effect on female rat VECs. Progesterone at 10(-9) and 10(-8) M had no effect on female rat VECs alone. When the ratio of E(2) to progesterone was 3/10, the proliferation of female rat VECs was accelerated (p < 0.05). When the ratio of E(2) to testosterone was 1/1, the proliferation of female rat VECs was also hastened (p < 0.05). However, when the ratio was reduced to 1/100, the hastening effect disappeared. CONCLUSION: Estrogen can speed up the proliferation of female and male rat VECs, while progesterone has no effect on female rat VECs alone. The balance of the ratio of E(2) to testosterone, E(2) to progesterone may play an important role in the proliferation of female rat VECs.