Objective To explore the effects of miR-373 and Janus kinase 2/signal transducer and activator of transcription 6 (JAK2/STAT6) signaling pathways on the M2 polarization of tumor associated macrophages (TAM) in rectal cancer. Methods THP-1 cells were induced into M0/M1/M2 macrophages, M0 macrophages were cocultured with Caco-2 cells to obtain TAM, Flow cytometry was used to detect the expression of CD86 and CD206, Real-time quantitative qPCR and Western blot were used to detect miR-373, inducible nitric oxide synthase (iNOS), toll-like receptor 4 (TLR-4), interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), arginase 1 (Arg1), chitinase 3-like 1 (Ym1), resistin like α (Fizz1), IL-10 mRNA and protein levels. TAM were transfected and divided into overexpressing miR-373 group (miR-373-TAM) and control group (miR-NC-TAM), overexpressing miR-373+JAK2-TAM group (miR-373 combined with JAK2-TAM) and control group (miR-373 combined with NC-TAM), and then cocultured with Caco-2 cells. Flow cytometry was used to detect the expression of CD206 in TAM; Real-time quantitative PCR and Western blot were used to detect miR-373, Arg1, Ym1, Fizz1, IL-10, JAK2, STAT6 mRNA and protein levels in TAM; CCK-8 assay, colony formation assay, and Transwell assay were used to detect the proliferation, migration, and invasion ability of Caco-2 cells. Thirty nude mice were randomly divided into Caco-2 cells group, Caco-2 cells combined with miR-NC-TAM group, and Caco-2 cells combined with miR-373-TAM group, with 10 mice in each group. Rats in each group were subcutaneously injected with pure Caco-2 cells, Caco-2 cells combined with TAM, and Caco-2 cells combined with TAM overexpressing miR-373. After 4 weeks of cell inoculation, immunofluorescence staining was used to detect F4/80⁺CD206⁺cells level in tumor tissue; Real-time quantitative PCR and Western blot were used to detect miR-373, JAK2, STAT6, Arg1, Ym1, Fizz1, IL-10 mRNA and protein levels in tumor tissues. Results TAM tended to M2 polarization. After overexpression of miR-373, miR-373 level in TAM was increased, while Arg1, Ym1, Fizz1, IL-10, JAK2, STAT6 mRNA and protein levels were decreased, proliferation, migration, invasion ability of Caco-2 cells were decreased; Overexpression of JAK2 could partially reverse the effect of overexpression of miR-373 on the M2 polarization of TAM and proliferation, migration, invasion ability of Caco-2 cells. TAM could promote tumor growth; Overexpression of miR-373 could inhibit tumor growth and inhibit M2 polarization of TAM. Conclusion miR-373 could inhibit M2 polarization of TAM in rectal cancer, and miR-373 might inhibit proliferation and metastasis of rectal cancer cells by regulating the JAK2/STAT6 pathway.

2020-02-01·Shanghai kou qiang yi xue = Shanghai journal of stomatology

[Effect of mi-138 targeting PLD2 gene on proliferation and migration of oral cancer cells].

Article

作者: Ding, Xu ; Ye, Jin-Hai ; Wu, He-Ming ; Li, Huai-Qi

PURPOSE:

To investigate the effect of miR-138 targeting PLD2 gene on proliferation and migration of oral cancer cells.

METHODS:

After oral cancer cells were transfected with miR-138, the expression level of microRNA-138 was detected by RT-PCR assay, the proliferation ability was detected by MTT assay, and cell cycle distribution was detected by flow cytometry. Transwell migration assay was used to detect cell migration ability, Western blotting assay was used to detect the expression levels of MMP-9, PLD2 and cyclin D1 in gastric cancer cells. Luciferase assay was used to report the targeting relationship between microRNA-138 and PLD2 gene. SPSS 21.0 software package was used to analyze the data.

RESULTS:

After miR-138 was transfected into oral cancer cells, the relative expression level of miR-138 was 4.28±0.16, which was significantly higher than that of blank control and miR-NC group (P<0.05). Luciferase reporter gene assay showed that the relative activity of PLD2 wild plasmid luciferase was significantly lower in oral cancer cells transfected with microRNA-138 than in other groups (P<0.05); The expression level of PLD2 gene in miR-138 group was significantly lower than that in blank control group and miR-NC group (P<0.05). After oral cancer cells were transfected with miR-138, the proliferation ability of oral cancer cells in miR-373 group was significantly lower than that in control group and miR-NC group (P<0.05). Flow cytometry showed that the ratio of G0/G1 phase was (64.39±6.49)% in the group of miR-138, which was significantly higher than that in the blank control group and the group of miR-NC(P<0.05); The ratio of S phase in the group of miR-138 was(13.28±3.16)%, which was significantly lower than that in the blank control group and the group of miR-NC (P<0.05); There was no significant difference in the ratio of G2/M phase among the groups (P>0.05). Transwell experiment showed that the number of migrating cells transfected with miR-138 in oral cancer cells was 138.46±24.37, which was significantly lower than that in blank control group and miR-NC group(P<0.05). Western blotting experiments showed that the relative levels of MMP-9, vimentin and cyclin D1 in the miR-138 group were 0.14±0.04, 0.17±0.02 and 0.15±0.03, respectively, which were significantly lower than those in the blank control group and the miR-NC group(P<0.05).

CONCLUSIONS:

miR-138 can target PLD2 gene expression and inhibit the proliferation and migration of oral cancer cells.

2019-01-01·International journal of clinical and experimental pathology

miR-373 inhibits nasopharyngeal carcinoma cell migration and invasion by targeting MARCH5.

Article

作者: Song, Xiang ; Li, Xiao ; Wang, Jia ; Zhang, Shengxiao ; Sun, Hehua ; Zhang, Yue ; Huang, Zehao

The abnormal expression of microRNAs (miRNAs) is critical for the development of human cancers. However, the functions of many miRNAs remain to be elucidated. miR-373 was reported to involve the tumorigenesis of multiple cancers, but its role in nasopharyngeal carcinoma (NPC) is not clear. Quantitative real-time PCR was performed to analyze miR-373 expression in NPC cell lines. The connection between membrane associated ring-CH-type finger 5 (MARCH5) and miR-373 was analyzed using a luciferase activity reporter assay and western blot. A cell counting kit-8 assay, a colony formation assay, and a wound-healing assay were performed to investigate the biological functions of miR-373 and MARCH5. We showed miR-373 expression is downregulated, and MARCH5 expression is upregulated, in NPC cells. MARCH5 was validated as a direct target of miR-373. miR-373 regulates NPC cell proliferation, colony formation, and cell migration by regulating MARCH5. In conclusion, our study showed that miR-373 has a tumor suppressive role in NPC by targeting MARCH5. This may provide novel therapeutic targets for NPC.

100 项与 MIR-373 相关的药物交易

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