Chong-Zhen Qin

A Long Noncoding RNA ZEB1-AS1 Promotes Tumorigenesis and Predicts Poor Prognosis in Glioma

Emerging studies show that long noncoding RNAs (lncRNAs) have important roles in carcinogenesis. lncRNA ZEB1 antisense 1 (ZEB1-AS1) is a novel lncRNA, whose clinical significance, biological function, and underlying mechanism remains unclear in glioma. Here, we found that ZEB1-AS1 was highly expressed in glioma tissues, being closely related to clinical stage of glioma. Moreover, patients with high ZEB1-AS1 levels had poor prognoses, with the evidence provided by multivariate Cox regression analysis indicating that ZEB1-AS1 expression could serve as an independent prognostic factor in glioma patients. Functionally, silencing of ZEB1-AS1 could significantly inhibit cell proliferation, migration, and invasion, as well as promote apoptosis. Knockdown of ZEB1-AS1 significantly induced the G0/G1 phase arrest and correspondingly decreased the percentage of S phase cells. Further analysis indicated that ZEB1-AS1 could regulate the cell cycle by inhibiting the expression of G1/S transition key regulators, such as Cyclin D1 and CDK2. Furthermore, ZEB1-AS1 functioned as an important regulator of migration and invasion via activating epithelial to mesenchymal transition (EMT) through up-regulating the expression of ZEB1, MMP2, MMP9, N-cadherin, and Integrin-β1 as well as decreasing E-cadherin levels in the metastatic progression of glioma. Additionally, forced down-regulation of ZEB1-AS1 could dramatically promote apoptosis by increasing the expression level of Bax and reducing Bcl-2 expression in glioma. Taken together, our data suggest that ZEB1-AS1 may serve as a new prognostic biomarker and therapeutic target of glioma.

HLA-B*58:01 is strongly associated with allopurinol-induced severe cutaneous adverse reactions in Han Chinese patients: a multicentre retrospective case-control clinical study.

DEAR EDITOR, Severe cutaneous adverse drug reactions (SCARs) are rare but life-threatening conditions, which can even be fatal in some patients. These include drug reaction with eosinophilia and systemic symptoms (DRESS), Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). Over one hundred drugs have been associated with SCARs, one of which is allopurinol, a drug widely used to treat hyperuricaemia and its complications. Allopurinol-induced DRESS/SJS/TEN has previously been shown to occur almost exclusively in individuals positive for the human leucocyte antigen allele HLA-B*58:01 and of Han Chinese or Thai descent; this variant allele is commonly found in these specific Asian subpopulations. However, in Japanese and white populations, where the presence of HLAB*58:01 is less frequent, only a moderate association between allopurinol-induced SCARs and HLA-B*58:01 has been seen. To elucidate this relationship we conducted a multicentre retrospective case–control study of Han Chinese patients. This study was registered in the Chinese Clinical Trial Registry with number ChiCTR-RCC-12002927 and was approved by the independent ethics committee of the Institute of Clinical Pharmacology, Central South University (CTXY-1100112). Each participant was subjected to HLA-B sequencing to guarantee an accurate genotype. In total 92 patients with allopurinol-induced SCARs, 75 allopurinol-tolerant patients and 99 healthy volunteers from 19 centres across China were enrolled in this study. The inclusion criteria were as follows. All participants had to be of Han Chinese descent. Patients with allopurinolinduced SCARs were diagnosed with DRESS, SJS, SJS/TEN overlap or TEN by dermatologists according to the RegiSCAR or Roujeau criteria. SCARs had to occur within 3 months of allopurinol use, with diminished or relieved symptoms upon withdrawal. DRESS was defined according to the RegiSCAR criteria: rash in combination with fever, lymphadenopathy and haematological abnormalities (e.g. eosinophilia, atypical lymphocytes, leucocytosis, thrombocytopenia), along with involvement of at least one internal organ (e.g. hepatitis, nephritis). SJS and TEN are more severe reactions and commonly overlap in the clinic. SJS/TEN was characterized by fever and mucocutaneous lesions (mouth, lips, genital and anal regions), which led to epidermal death and sloughing. SJS was defined as skin detachment < 10% of the total body surface area, SJS/TEN overlap as 10–30% and TEN as > 30% skin detachment. Allopurinol-tolerant individuals were defined as patients using allopurinol for at least 3 months without any evidence of cutaneous adverse events. Healthy volunteers were Han Chinese individuals without clinical diseases in their medical history, and who had never received allopurinol treatment. The exclusion criteria were patients with a medical history of bone marrow transplantation, chemotherapy or cancer. Genomic DNA was extracted from the study participants’ blood samples for HLA-B genotyping by sequence-based typing. The odds ratio (OR), 95% confidence interval (CI), Pvalue, corrected P-value (Pc), positive predictive value and negative predictive value were calculated. The Pc-values were adjusted using Bonferroni’s post hoc correction for multiple comparisons to account for the observed alleles (48 HLA-B alleles were identified in three groups). The Pc-values were therefore multiplied by a factor of 48. The demographic characteristics of the participants are listed in Table 1. The clinical features of a patient with TEN are presented in Figure 1a. In the allopurinol-induced SCAR group, the subgroups of patients with DRESS, SJS, SJS/TEN overlap and TEN contained 41 (44%), 33 (36%), seven (8%) and 11 (12%) patients, respectively (Fig. 1b). Higher prevalences of allopurinol-induced SCARs (65%) and allopurinol tolerance (95%) were seen in male patients than in female patients (Fig. 1c). This can be explained by the high incidence of gout and hyperuricaemia in elderly men. The age distribution was about equal between groups (Fig. 1c). The group with allopurinol-induced SCARs received a shorter period of treatment with allopurinol, varying from 1 day to 64 days (mean 22 1), compared with 98–8760 days (mean 1153 303) in the allopurinol-tolerant group. Some patients exhibited higher sensitivity to allopurinol and had skin eruptions at day 1. Two patients experienced a second SCAR due to re-exposure to allopurinol (patients 3 and 81, Table S1; see Supporting Information). One patient had multidrug sensitivity (patient 3, Table S1). The mean given dose of allopurinol in the SCAR group was lower than in the tolerant group (186 5 vs. 289 8 mg), because most of the patients with SCARs had insufficient renal function. In terms of additional medications, 81% (57 of 70) of the patients used other drugs together with

Comparison of HPV genotyping and methylated ZNF582 as triage for women with equivocal liquid-based cytology results

IntroductionThe interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). Recently, methylated zinc finger protein 582 (ZNF582) (ZNF582m) was reported to be highly associated with cervical cancer. In this study, we compared the performance of ZNF582m detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H).Case descriptionTwo hundred and forty-two subjects with equivocal papanicolaou smear (Pap smear) results were recruited in this hospital-based and case-controlled study. The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and then for ZNF582m and HPV-HR detection. The level of ZNF582m was quantified by real-time methylation-specific PCR after bisulfite conversion. The HPV-HR test was performed by using a nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and HPV type-specific primers.Discussion and evaluationSignificant associations were observed between ZNF582m and the risk of cervical intraepithelial neoplasia grade 3 or higher (CIN3+; odds ratio = 15.52, 95% confidence interval (CI): 7.73 to 31.18). The sensitivity and specificity of ZNF582m for women with CIN3+ were 82.43% and 76.79%, respectively. High sensitivity (99.33%) but low specificity (38.76%) was observed for HPV-HR. When combining both positive results of ZNF582m and HPV-HR, the sensitivity and specificity were 82.43% and 81.55%, respectively. The sensitivity and specificity of ZNF582m or HPV-16/18 were 89.19% and 70.24%, respectively. However, the sensitivity and specificity of ZNF582m combined with HPV-16/18 (both ZNF582m and HPV-16/18 positive results) were 59.46% and 94.64%, respectively.ConclusionsZNF582m provides a promising triage tool for women with ASC.To effectively manage ASC patients, a new strategy co-testing for ZNF582m and HPV-16/18 genotyping was proposed. This strategy could reduce the number of patients referred for colposcopic examination and thus provide a feasible follow-up solution in the regions where colposcopy is not readily available. This strategy could also prevent women from experiencing unnecessary anxiety caused by HPV-HR.

In Vitro and in Vivo Inhibitory Effects of Glycyrrhetinic Acid in Mice and Human Cytochrome P450 3A4

Glycyrrhetinic acid (GA) has been used clinically in the treatment of patients with chronic hepatitis. This study evaluated the effect of GA on the activity of five P450(CYP450) cytochrome enzymes: CYP2A6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4, in human liver microsomes (HLMs) and recombinant cDNA-expressed enzyme systems using a HPLC-MS/MS CYP-specific probe substrate assay. With midazolam as the probe substrate, GA greatly decreased CYP3A4 activity with IC50 values of 8.195 μM in HLMs and 7.498 μM in the recombinant cDNA-expressed CYP3A4 enzyme system, respectively. It significantly decreased CYP3A4 activity in a dose- but not time-dependent manner. Results from Lineweaver–Burk plots showed that GA could inhibit CYP3A4 activity competitively, with a Ki value of 1.57 μM in HLMs. Moreover, CYP2C9 and CYP2C19 could also be inhibited significantly by GA with IC50 of 42.89 and 40.26 μM in HLMs, respectively. Other CYP450 isoforms were not markedly affected by GA. The inhibition was also confirmed by an in vivo study of mice. In addition, it was observed that mRNA expressions of the Cyps2c and 3a family decreased significantly in the livers of mice treated with GA. In conclusion, this study indicates that GA may exert herb-drug interactions by competitively inhibiting CYP3A4.

Clinical Significance of Long Non-Coding RNA CASC8 rs10505477 Polymorphism in Lung Cancer Susceptibility, Platinum-Based Chemotherapy Response, and Toxicity

Long non-coding RNA (lncRNA) CASC8 rs10505477 polymorphism has been identified to be related to risk of many kinds of cancers, such as colorectal cancer, gastric cancer, and invasive ovarian cancer, and it may be involved in the prognosis of gastric cancer patients who have received platinum-based chemotherapy after surgical treatment. So far, there is no study investigating the clinical significance of lncRNA CASC8 rs10505477 in lung cancer susceptibility and treatment. In this study, we genotyped 498 lung cancer patients and 213 healthy control subjects to explore the correlation between the rs10505477 polymorphism and lung cancer risk in a Chinese population. Among the 498 patients, 467 were selected for the chemotherapy response and toxicity study. We found that the single nucleotide polymorphisms (SNP) rs10505477 was greatly related to lung cancer risk in male and adenocarcinoma subgroups in recessive model (adjusted OR = 0.51, 95%CI = 0.29–0.90, p = 0.02; adjusted OR = 0.52, 95%CI = 0.30–0.89, p = 0.02, respectively). It was also closely correlated with platinum-based chemotherapy response in dominant model (adjusted OR = 1.58, 95%CI = 1.05–2.39, p = 0.03). Additionally, we observed that CASC8 rs10505477 polymorphism was significantly relevant to severe hematologic toxicity in non-small-cell lung cancer (NSCLC) subgroup in dominant model (adjusted OR = 0.59, 95%CI = 0.35–0.98, p = 0.04) and in additive model (adjusted OR = 0.62, 95%CI = 0.43–0.90, p = 0.01). Furthermore, it was found that rs10505477 polymorphism was greatly associated with gastrointestinal toxicity in SCLC and cisplatin subgroups in dominant model (adjusted OR = 7.82, 95%CI = 1.36–45.07, p = 0.02; adjusted OR = 1.94, 95%CI = 1.07–3.53, p = 0.03, respectively). Thus, lncRNA CASC8 rs10505477 could serve as a possible risk marker for diagnosing lung cancer, and could be used to forecast the response and toxicity of platinum-based treatment in lung cancer patients.

Mechanism-based inhibition of Alantolactone on human cytochrome P450 3A4 in vitro and activity of hepatic cytochrome P450 in mice

ETHNOPHARMACOLOGICAL RELEVANCE Alantolactone (AL), one of the main active ingredients in Inula helenium L., has been included in various prescriptions of traditional Chinese medicine. The effects of AL on cytochrome P450 (CYP450) were still unclear. This study evaluated the inhibitory effect of AL on cytochrome P450s in vitro and in vivo. MATERIALS AND METHODS The inhibitory effects of AL on the CYPs activity were evaluated in human liver microsomes (HLMs) and recombinant cDNA-expressed enzymes incubation system, and then determined by LC-MS/MS based CYPs probe substrate assay. C57BL/6 mice were treated AL orally (0, 25, 50, 100 mg/kg) for 15 days. The inhibitory effects of AL on major Cyps in mice were examined at both the mRNA and enzyme activity levels. RESULTS AL showed a potent inhibitory effect on CYP3A4 activity with IC50 values of 3.599 (HLMs) and 3.90 (recombinant CYP3A4) μM, respectively. AL strongly decreased CYP3A4 activity in a dose-dependent but not time-dependent way in HLMs. Results from typical Lineweaver-Burk plots showed that AL could inhibit CYP3A4 activity noncompetitively, with a Ki value of 1.09 μM in HLMs. Moreover, activity of CYP2C19 could also be inhibited by AL with IC50 of 36.82 μM. Other CYP450 isoforms were not markedly affected by AL. The inhibition was also validated by in vivo study of mice. AL significantly decreased mRNA expression of Cyp2c and 3a family. CONCLUSION The study indicates that herb-drug interaction should be paid more attention between AL and drugs metabolized by CYP3A4.

Advances in Molecular Signaling Mechanisms of β-Phenethyl Isothiocyanate Antitumor Effects

β-Phenethyl isothiocyanate (PEITC) is an important phytochemical from cruciferous vegetables and is being evaluated for chemotherapeutic activity in early phase clinical trials. Moreover, studies in cell culture and in animals found that the anticarcinogenic activities of PEITC involved all the major stages of tumor growth: initiation, promotion, and progression. A number of mechanisms have been proposed for the chemopreventive activities of this compound. Here, we focus on the major molecular signaling pathways for the anticancer activities of PEITC. These include (1) activation of apoptosis pathways; (2) induction of cell cycle arrest; and (3) inhibition of the survival pathways. Furthermore, we also discussed the regulation of drug-metabolizing enzymes, including cytochrome P450s, metabolizing enzymes, and multidrug resistance.

Progesterone Prevents High-Grade Serous Ovarian Cancer by Inducing Necroptosis of p53-Defective Fallopian Tube Epithelial Cells

High-grade serous ovarian carcinoma (HGSOC) originates mainly from the fallopian tube (FT) epithelium and always carries early TP53 mutations. We previously reported that tumors initiate in the FT fimbria epithelium because of apoptotic failure and the expansion of cells with DNA double-strand breaks (DSB) caused by bathing of the FT epithelial cells in reactive oxygen species (ROSs) and hemoglobin-rich follicular fluid (FF) after ovulation. Because ovulation is frequent and HGSOC is rare, we hypothesized that luteal-phase progesterone (P4) could eliminate p53-defective FT cells. Here we show that P4, via P4 receptors (PRs), induces necroptosis in Trp53-/- mouse oviduct epithelium and in immortalized human p53-defective fimbrial epithelium through the TNF-α/RIPK1/RIPK3/MLKL pathway. Necroptosis occurs specifically at diestrus, recovers at the proestrus phase of the estrus cycle, and can be augmented with P4 supplementation. These results reveal the mechanism of the well-known ability of progesterone to prevent ovarian cancer.

Genetic variation of CYP3A5 influences paclitaxel/carboplatin‐induced toxicity in Chinese epithelial ovarian cancer patients

Combination chemotherapy with platinum and taxane is the first‐line treatment for ovarian cancer. The dose‐limiting toxicities of these drugs include neuropathy, leukopenia, and neutropenia, but they exhibit substantial interindividual variability. This study investigated the relationship between CYP3A5 polymorphisms and paclitaxel/carboplatin‐induced toxicity in Chinese epithelial ovarian cancer patients. Seventy‐five patients with epithelial ovarian cancer were recruited. After combination chemotherapy, genotype analysis was conducted, and toxic effects were evaluated according to the Common Toxicity Criteria. A significant association was found between myelosuppression and the CYP3A5*3 genotype. CYP3A5*3/*1 patients showed a significantly higher risk of developing leukopenia (P < .001; Pearsons χ2 test) and neutropenia (P < .001; Pearsons χ2 test) than CYP3A5*3*3 patients. CYP3A5*3/*3 patients had significantly higher median leukocyte and neutrophil nadir counts than CYP3A5*3*1 patients (P < .001, Mann–Whitney U test). However, we did not observe an association between neuropathy and CYP3A5*3 in this study (P =.64; Pearsons χ2 test). This is the first study to verify the influence of CYP3A5 polymorphisms on paclitaxel/carboplatin‐induced toxicity in Chinese epithelial ovarian cancer patients. Our findings suggest that interindividual variability in paclitaxel/carboplatin‐induced myelosuppression can be predicted by CYP3A5*3 genotyping and that incorporation of CYP3A5*3 genetic data in treatment selection could help to reduce myelosuppression events, thereby individualizing paclitaxel/carboplatin pharmacotherapy.

Downregulation of MicroRNA-320d predicts poor overall survival and promotes the growth and invasive abilities in glioma.

Previous studies have demonstrated that miRNAs play an important role in tumor development and progression. The role of miR‐320d has been studied in several cancers except for glioma. The aim of the study was to investigate the expression levels, biological function, and mechanism of miR‐320d in glioma. The expression levels of miR‐320d were detected in glioma tissues and cell lines (U87 and U251) by RT‐qPCR. Cell proliferation, colony formation, apoptosis, cell cycle, and transwell assays were performed in glioma cell lines transfected with miR‐320d mimics or controls to evaluate the effects of miR‐320d in vitro. The expression levels of invasive‐related proteins were determined by Western blot analysis. Results showed that the expression of miR‐320d was significantly decreased in glioma tissues and cell lines. Overexpression of miR‐320d could significantly suppress cell growth, migration and invasion, and induced cell apoptosis as well as cell cycle at G0/G1 arrest in U87 and U251 cell lines. Additionally, expression levels of MMP‐2, MMP‐9, N‐cadherin, and integrin‐β1 reduced, while E‐cadherin increased in miR‐320d mimic group. Overall, this study is the first to demonstrate that miR‐320d may serve as an independent prognostic factor, indicating that miR‐320d is a biomarker for prognosis and therapy in glioma.