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CPT-11 is one of the drugs employed in colorectal cancer treatment and has faced challenges in the form of resistance. The insulin-like growth factor 1 receptor is a tyrosine kinase receptor that mediates cancer cell survival and drug resistance. It is frequently overexpressed in colorectal cancer and has previously been identified as a microRNA target. MicroRNAs are non-coding RNA molecules that regulate gene function by suppressing messenger RNA translation. Studies have demonstrated that natural compounds can regulate microRNA function and their target genes. Therefore, combining natural compounds with existing cancer drugs can enhance the therapeutic efficacy. We investigated a natural compound, Aloin, for the potential sensitization of colorectal cancer to CPT-11. We used western blot, MTT cell viability assay, flow cytometry, and microRNA/gene knockdown and overexpression experiments, as well as an in vivo mouse model. Our investigation revealed that combining Aloin with CPT-11 exerts an enhanced anti-tumor effect in colorectal cancer. This combination reduced cell viability and induced apoptosis, both in vivo and in vitro. Furthermore, this combination upregulated miRNA-133b, while downregulating the IGF1R and its downstream MEK/ERK, and PI3K/AKT/mTOR pathways. Our findings suggests that CPT-11 and Aloin are potential combination treatment partners against colorectal cancer. MicroRNA-133b may serve as a co-therapeutic target with IGF1R against colorectal cancer, which might overcome the existing treatment limitations.

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Recombinant adeno-associated virus (rAAV) is an extremely attractive vector in the in vivo delivery of gene therapy as it is safe and its genome is simple. However, challenges including low permissiveness to specific cells and restricted tissue specificity have hindered its clinical application. Based on the previous studies, epidermal growth factor receptor-protein tyrosine kinase (EGFR-PTK) negatively regulated rAAV transduction, and EGFR-positive cells were hardly permissive to rAAV transduction. We constructed a novel rAAV-miRNA133b vector, which co-expressed miRNA133b and transgene, and investigated its in vivo and in vitro transduction efficiency. Confocal microscopy, live-cell imaging, pharmacological reagents and labelled virion tracking were used to analyse the effect of miRNA133b on rAAV2 transduction and the underlying mechanisms. The results demonstrated that miRNA133b could promote rAAV2 transduction and the effects were limited to EGFR-positive cells. The increased transduction was found to be a direct result of decreased rAAV particles degradation in the cytoplasm and enhanced second-strand synthesis. ss-rAAV2-miRNA133b vector specifically increased rAAV2 transduction in EGFR-positive cells or tissues, while ss-rAAV2-Fluc-miRNA133b exerted an antitumor effect. rAAV-miRNA133b vector might emerge as a promising platform for delivering various transgene to treat EGFR-positive cell-related diseases, such as non-small-cell lung cancer.

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Objective:To investigate the expressions of miRNA-133b (miR-133b) and miRNA-150 (miR-150) in non-small cell lung cancer (NSCLC) and their clinical significances.Methods:Thirty patients with NSCLC who underwent surgery and were pathologically diagnosed in the First People's Hospital of Lianyungang from June 2018 to June 2019 were selected. Before surgery, 10 ml fasting peripheral blood of patients was collected in the morning. The fresh cancer tissues, adjacent tissues (distance from cancer tissues ≤ 3 cm), normal lung tissues (distance from cancer tissues > 5 cm) were collected after surgery. Thirty people who had a physical examination during the same period were served as the healthy control group, and 10 ml fasting peripheral blood was collected in the morning. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expressions of miR-133b and miR-150 in the serum of the two groups of subjects and the tissues of NSCLC patients. The relationships between serum miR-133b and miR-150 levels and the clinical features in NSCLC patients were analyzed. The pathological diagnosis was used as the gold standard, and the receiver operating characteristic (ROC) curve was used to determine the efficacy of miR-133b and miR-150 in diagnosing NSCLC.Results:In NSCLC patients, the relative expressions of miR-133b in cancer tissues, adjacent tissues and normal lung tissues were 0.55±0.17, 0.68±0.11 and 0.69±0.12, and the expression of miR-133b in cancer tissues was lower than that in adjacent tissues and normal lung tissues (both P < 0.01). The relative expressions of miR-150 in cancer tissues, adjacent tissues and normal lung tissues were 1.19±0.14, 1.13±0.13 and 1.09±0.12, and there was no significant difference in the expression of miR-150 between cancer tissues and adjacent tissues ( t = 1.98, P = 0.051), and the expression of miR-150 in cancer tissues was higher than that in normal lung tissues ( t = 3.06, P = 0.003). The relative expression of plasma miR-133b in NSCLC patients was lower than that in healthy controls (0.43±0.11 vs. 0.55±0.16, t = 3.68, P<0.05); the relative expression of plasma miR-150 was higher than that in healthy controls (1.14±0.13 vs. 1.04±0.12, t = -3.18, P < 0.05). The expression level of miR-133b in the serum of NSCLC patients was related to lymph node metastasis ( P = 0.003), but not related to the patients' gender, age, pathological type, and TNM stage (all P > 0.05). The expression level of miR-150 in serum of NSCLC patients was not related to the clinicopathological characteristics (all P > 0.05). According to the ROC curve, the area under the curve of miR-133b and miR-150 in serum of NSCLC patients were 0.732 and 0.726, and the area under the curve of the combined detection of the two was 0.811. Conclusions:The expression levels of miR-133b in cancer tissues and serum of NSCLC patients decrease, and the expression levels of miR-150 increase. The expression trends of the two in cancer tissues and serum of NSCLC patients are consistent. miR-133b may be related to the malignant degree, invasion and metastasis of NSCLC. miR-133b and miR-150 may become molecular markers for the diagnosis of NSCLC, and the combined detection of the two has higher diagnostic efficiency.

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BACKGROUND: Splicing factor 3b subunit 4 (SF3B4) is a splicing factor and potential oncogene in hepatocellular carcinoma (HCC); however, its regulatory mechanism is yet unclear. We aimed to determine the role of SF3B4 in HCC and the underlying mechanism. METHODS: To investigate the association between alternative splicing events and miRNAs, putative miRNAs were screened using TargetScan. Expression levels of and prognostic information for SF3B4 and miRNAs were determined based on public genomic data and clinical samples. Then, we examined the possible roles of SF3B4 and miRNA-133b in HCC cells and a xenograft mouse model. Pearson correlation analysis and in vitro experiments verified SF3B4 as a miRNA-133b target. Protein levels of key targets from the SF3B4 signaling pathway were estimated using western blotting. FINDINGS: The expression of SF3B4 was upregulated in HCC tissues and cell lines whereas, the expression of miRNA-133b was downregulated. MiRNA-133b negatively regulated the expression of SF3B4. Effects of SF3B4 overexpression were partially abolished by miRNA-133b mimics, confirming that SF3B4 is a target of miRNA-133b. Moreover, molecules associated with SF3B4, including KLF4, KIP1, and SNAI2, were also modulated by miRNA-133b. INTERPRETATION: SF3B4 plays a crucial role in HCC and is negatively regulated by miRNA-133b. The miRNA-133b/ SF3B4 axis may serve as a new therapeutic target for HCC treatment. FUND: China National Funds for Distinguished Young Scientists (No.81425019), the State Key Program of National Natural Science Foundation of China (No.81730076), Shanghai Science and Technology Committee Program (No.18XD1405300) and Specially-Appointed Professor Fund of Shanghai (GZ2015009). China National Funds for National Natural Science Fund (No.81672899).

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Objective To explore the diagnostic value of the quantitative detection of plasma miRNA-125b and miRNA-133b in children with asthma.Methods Thirty asthmatic patients were enrolled in this study and collected the blood specimens during acute phase and stable phase respectively (AP group and SP group).Thirty allergic rhinitis children (AR group) and thirty healthy children were recruited to the control group (NC group).The levels of miRNA in different groups were detected by qRT-PCR.The performance of miRNA-125b and miRNA-133b were evaluated by receiver operating characteristic curves (ROC) and the area under the curve (AUC) (95％CI).Results The relative expression of miRNA-125b in AP group and SP group were significantly higher than A R group (t=3.913,3.120,P＜0.01),miRNA-133b.In AP group and AR group the expression of miRNA-133b were significantly higher than control group (t=4.426,4.720,P＜0.01).The detection of miRNA-125b yielded an area under the curve of ROC of 0.7989,(95 ％ CI:0.7111～ 0.8864) in discriminating asthmatic patients from healthy group.And the miRNA-133b was 0.7274 (95％CI:0.586 5～0.863 0) in discriminating asthmatic patients during the acute phase from healthy group.Conclusion The relative expression of miRNA-125b in children with asthma was significantly higher than that in AR group and NC group,miRNA-125b may prove to be a non-invasive biomarker for the auxiliary diagnosis of asthma especially when the relative expression up to 1.998.

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Sertoli cells play critical roles in regulating spermatogenesis and they can be reprogrammed to the cells of other lineages, highlighting that they have significant applications in reproductive and regenerative medicine. The fate determinations of Sertoli cells are regulated precisely by epigenetic factors. However, the expression, roles, and targets of microRNA (miRNA) in human Sertoli cells remain unknown. Here we have for the first time revealed that 174 miRNAs were distinctly expressed in human Sertoli cells between Sertoli-cell-only syndrome (SCOS) patients and obstructive azoospermia (OA) patients with normal spermatogenesis using miRNA microarrays and real time PCR, suggesting that these miRNAs may be associated with the pathogenesis of SCOS. MiR-133b is upregulated in Sertoli cells of SCOS patients compared to OA patients. Proliferation assays with miRNA mimics and inhibitors showed that miR-133b enhanced the proliferation of human Sertoli cells. Moreover, we demonstrated that GLI3 was a direct target of miR-133b and the expression of Cyclin B1 and Cyclin D1 was enhanced by miR-133b mimics but decreased by its inhibitors. Gene silencing of GLI3 using RNA inference stimulated the growth of human Sertoli cells. Collectively, miR-133b promoted the proliferation of human Sertoli cells by targeting GLI3. This study thus sheds novel insights into epigenetic regulation of human Sertoli cells and the etiology of azoospermia and offers new targets for treating male infertility.

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