Expression of HMGA1 in uveal melanoma and its effect on cell proliferation and invasion / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the expression of high mobility group A1(HMGA1)protein in uveal melanoma(UM)tissues, and the effect of inhibiting the expression of <i>HMGA1</i> on cell proliferation and invasion. <p>METHODS: A total of 53 cases(53 eyes)of UM patients who underwent surgical treatment in our hospital from February 2014 to August 2019 were selected. In the same period, 34 cases(34 eyes)of normal uveal tissues removed from the eye due to trauma were selected. The expression of HMGA1 protein in tissues was detected by immunohistochemistry. The human UM cell line M23 was cultured and divided into <i>HMGA1</i> downregulation group, negative control group and blank group, respectively, transfected with <i>HMGA1</i> interference sequence, negative control sequence and without any treatment. The expression of <i>HMGA1 </i>was detected by real-time quantitative PCR. The cell proliferation ability was detected by CCK-8 method, and the cell migration and invasion abilities were detected by Transwell method.<p>RESULTS: The positive expression rate of HMGA1 protein in UM tissue was 77%, which was higher than that in the normal uveal tissue, which was 29%(<i>P</i><0.001). Compared with the no scleral occurring infiltration, no ciliary body involving, and no extraocular growth, the positive expression rates of HMGA1 proteins in the scleral infiltration, ciliary body involving, and extraocular growth occurring tissues were increased(all <i>P</i><0.05). The relative expression level of <i>HMGA1</i> mRNA in cells in the <i>HMGA1</i> downregulation group was lower than that in the negative control group and the blank group. Compared with the negative control group and the blank group, the absorbance <i>OD</i> values of cells in the <i>HMGA1</i> downregulation group at 24, 48, 72 and 96h were decreased(<i>P</i><0.05). The number of migrating cells and the number of invading cells in the <i>HMGA1</i> downregulation group was significantly less than those in the negative control group and the blank group(<i>P</i><0.05). <p>CONCLUSION: The positive expression rate of HMGA1 protein in UM tissue is increased. Down regulation the expression of <i>HMGA1</i> in M23 cells can reduce cell proliferation and inhibit cell migration and invasion.

miR-424 modulates radio-sensitivity of breast cancer cells via targeting HMGA1 / 中国肿瘤生物治疗杂志

@# Objective: To explore the effect of miR-424/HMGA1 (high mobility proteinA1) axis on the radio-sensitivity of breast cancer cells and the possible mechanism. Methods:Atotal of 50 cases of breast cancer tissues from patients, who underwent surgical resection at the Department of Oncological Radiotherapy, Wuxi Fourth People’s Hospital from April 2014 to April 2017, were collected for this study. Real-time quantitative polymerase chain reaction (qPCR) and Western blotting were performed to evaluate the mRNA and protein expressions of miR-424 and HMGA1 in breast cancer tissues of radiation sensitive and insensitive patients. After being treated with different doses of 60Co γ-ray radiation (0, 2, 4, 6 and 8 Gy), the expression changes of miR-424 and HMGA1 in breast cancer MDA-MB-468 cells were observed. Subsequently, miR-424 mimic/inhibitor and pcDNA-HMGA1 were transfected into MDA-MB-468 cells, and the effect of miR-424 on cell proliferation, invasion and apoptosis of radiation-treated MDA-MB-468 cells were evaluated by colony formation assay, MTT assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Furthermore, dual luciferase reporter gene assay was used to verify whether HMGA1 was a target gene of miR-424. Results: The patients in radio-sensitive group exhibited higher miR-424 expression but lower HMGA1 expression than the patients in insensitive group (all P<0.01). Compared with the cells treated with 0, 2 and 4 Gy radiation, the cells treated with 6 and 8Gy radiation exhibited significantly higher apoptosis rate and miR-424 expression but lower HMGA1 expression and cell invasion (all P<0.01). Moreover, luciferase reporter gene assay confirmed that miR-424 down-regulated HMGA1 expression. Mechanistically, miR-424 significantly inhibited cell proliferation, invasion and induced apoptosis of MDA-MB-468 cells (all P<0.01) via targeted down-regulating HMGA1, and further upregulated the radio-sensitivity of breast cancer cells. Conclusion: miR-424/HMGA1 axis regulates the radio-sensitivity of breast cancer, and over-expression of miR-424 may increase the sensitivity of MDA-MB-468 cells to γ-ray radiation therapy.

Expression and clinical significance of HMGA1 and CXCR4 in invasive ductal carcinoma of breast / 临床与实验病理学杂志

Purpose To investigate the expression of high mobility group protein Al (HMGA1) and C-X-C chemokine receptor 4 (CXCR4) in breast invasive ductal carcinoma and its clinical significance. Methods Immunohistochemical method was used to detect the expression of HMGA1 and CXCR4 in 105 cases of breast invasive ductal carcinoma and 80 cases of breast adenosis. The correlation between HMGA1 and CXCR4 expression and clinicopathological features was analyzed. Results The positive rate of HMGA1 and CXCR4 in breast invasive ductal carcinoma was significantly higher than that of breast adenosis(77.14％ vs 26.25％, 73.33％ vs 23.75％ ), the difference was statistically significant (P＜ 0.001). There was no significant correlation between HMGA1 and CXCR4 expression in breast cancer tissues (r = 0.104, P =0.289), suggesting that the expression of them were independent of each other. The combined detection of HMGA1 and CXCR4 could improve the sensitivity of diagnosis of (either positive) and specificity of(both positive). The positive rate of CXCR4 in PR positive breast cancer (87.5％ ) was higher than that in PR negative(60.0％ ), the difference was statistically significant (P =0.008) Conclusion HMGA1 is highly expressed in breast invasive ductal carcinoma, and CXCR4 expression is mainly low in breast invasive ductal carcinoma. HMGA1 and CXCR4 have higher sensitivity, and the combined detection of them can significantly improve the sensitivity and specificity of breast cancer diagnosis. The high expression of HMGA1 and CXCR4 in breast cancer has a certain clinical significance for the diagnosis and prognosis of breast cancer, which is expected to provide a new theoretical basis for the diagnosis and treatment of clinical breast cancer.

MicroRNA-26a inhibits cell migration and invasion in cervical cancer through targeting high mobility group protein A1(HMGA1) / 中华微生物学和免疫学杂志

Objective To investigate the role of microRNA-26a (miR-26a) in cell migration and invasion in cervical cancer and its regulatory effects on high mobility group protein A1(HMGA1). Methods Both HeLa and SiHa cells were divided into four groups:miR-26a mimic group,mimic control group,miR-26a inhibitor group and inhibitor control group. MiR-26a expression was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Target genes for miR-26a-5p were predicted by bioinformatics and verified by dual-luciferase reporting system. HMGA1 expression was evaluated by Western blot and qRT-PCR. Cell migration and invasion were analyzed by in vitro assays. Results (1) HMGA1 was predicted as one of the potential targets of miR-26a by TargetScan. Results of dual-luciferase activity assay further con-firmed that HMGA1 was directly regulated by miR-26a. (2) Expression of miR-26a and HMGA1 at mRNA level was respectively enhanced and inhibited in miR-26a mimic-transfected HeLa as well as SiHa cells as compared with those in the corresponding mimic control groups (P<0.05). On the contrary, expression of miR-26a and HMGA1 at mRNA level was respectively reduced and increased in miR-26a inhibitor groups as compared with those in the corresponding inhibitor control groups(P<0.05). (3) Expression of HMGA1 in miR-26a mimic-transfected HeLa and SiHa cells was lower than that in the corresponding mimic control groups(P<0.05). But expression of HMGA1 in miR-26a inhibitor groups was higher than that in the corre-sponding inhibitor control groups(P<0.05). (4) Abilities of cell migration and invasion were suppressed in miR-26a mimic groups as compared with those in the corresponding mimic control groups,but were enhanced in miR-26a inhibitor groups as compared with those in the corresponding inhibitor control groups(P<0.05). Conclusion MiR-26a can inhibit the proliferation and invasion of HeLa and SiHa cells through targeting HMGA1,suggesting that miR-26a is closely related to cervical cancer and might be associated with chemo-therapy resistance in cervical cancer. This study might provide a new strategy for the prevention and treat-ment of cervical cancer.

Mechanism of Lipid Induced Insulin Resistance: An Overview

Type 2 diabetes (T2D) is rapidly spreading throughout the world. It's an insidious disease and still treated in an indirect manner without having specific drug target. In majority cases T2D is treated with drugs that address type 1 diabetes, majority of drugs aim to increase insulin release although the root cause for T2D is not the dearth of insulin release, it occurs in the later stage of disease development. T2D silently progressed in the patient; it begins with insulin resistance that takes place due to the loss of insulin sensitivity. Though insulin resistance is the centre of pathogenesis, our treatment of the disease has not yet addressed it. It is now a fact that insulin resistance is manifested by lipid and fatty acids (FAs) play a critical role in blunting insulin sensitivity. Our understanding is still poor in deciphering how lipid impose insulin insensitivity, majority of workers suggest it is because of insulin signaling defects which implements insulin function in inhibiting glucose to the cell from circulation. Number of long chain saturated FA has been shown to produce insulin signaling defects. However, we really need further investigation to find specific target(s) for FA induced damage. In addition to these information, a new dimension of T2D is getting attractive is fetuin-A/alpha2-Heremans-Schmid Glycoprotein, a secretary protein from liver. Its gene locus has been identified as T2D susceptible. Fetuin-A's excess expression occurs by FA and it disrupts adipocyte function. It has been shown to be associated with T2D especially in obesity. In this review, we briefly discuss the present status on the mechanistic understanding of lipid induced insulin resistance that leads to T2D. More we understand the mechanism; opportunity to fight the battle with T2D will be increasing. Since, this field is now vast; we covered a few major events.

Construction and identification of a lentiviral vector harboring RNAi sequence targeting the human high mobility group A1 gene / 医学研究生学报

Objective: To construct and identify a lentiviral vector harboring RNAi sequence targeting the human high mobility group A1(HMGA1) gene.Methods: The effective sequence of siRNA targeting the HMGA1 gene confirmed in our previous study,the complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pGCL-GFP vector diced by the restriction enzyme of HpaⅠ and XhoⅠ,which contained the U6 promoter and green fluorescent protein(GFP).The resulting lentiviral vector containing HMGA1 shRNA was named LV-sh HMGA1 and confirmed by PCR and DNA sequencing.A total of 293T cells were cotransfected with LV-sh HMGA1,pHelper 1.0 and pHelper 2.0.All the virus stocks were produced by Lipofectamine2000-mediated transfection.The titer of the virus was tested according to the expression level of GFP.Results: PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the human HMGA1 gene was successfully inserted into the lentiviral vector.The titer of the recombinant lentiviral vector was 5?107 TU/ml.Conclusion: The successful construction of the lentiviral vector of HMGA1 has prepared the ground for further studies on the functions of the HMGA1 gene with the RNAi technique.

Effects of HMGA1 silence by RNA interference on the apoptosis and cell cycle of hepatocarcinoma cells / 中国免疫学杂志

Objective:To develop HMGA1-silencing SMMC-7721 cell and investigate its impacts on apoptosis and cell cycle in tumor cells.Methods:Constructing HMGA1-silencing SMMC-7721 cell through G418 selection of RNAi plasmid transfected cells;surveying the apoptosis,prliferation and cell cycle of the tumor cells by FACS and MTT.Results:Stable HMGA1-silencing cells were obtained successfully.The apoptosis percentage in RNAi group(29.46?3.04%) was significantly higher than that in the negative control (1.96?0.76%) or control (2.04?0.70%)within each P

The study of malignant transformation of NIH3T3 cells induced by transfection of HMGA1 gene / 中国免疫学杂志

Objective:To establish a mouse fibroblastic cell line stably transfected with HMGA1 gene,and to use the cell line to investigate the role of HMGA1 in tumor development and progression.Methods:Eukaryotic expression vector pcDNA3.0-HMGA1 was transfected into NIH3T3 by lipofectamine-mediation.Stable transfectants were selected by G418.The expression of extraneous HMGA1 gene was analyzed by RT-PCR and DNA sequencing.Cell growth was measured through MTT and the cell cycle by FCM.Soft agar colony formation was employed to analyze the ability of anchorage-independent growth.The expression of the immune inhibition factors of VEGF and FasL mRNA was detected by RT-PCR.Results:NIH3T3 cell lines stably expressing HMGA1 gene were successfully established.Comparing with controls,the HMGA1 gene-transfected cells grew faster,acquired the ability of anchorage-independent growth and expressed the immune inhibition factors of VEGF and FasL mRNA.Conclusion:Extraneous expression of HMGA1 gene can induce malignant transformation of NIH3T3 and modulate mRNA expression of immune inhibition factors.HMGA1 gene plays a key role in tumor development,progression and immune escape.