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Objective To investigate the expression of microRNA (miR)-513c-5p in cervical cancer and the mechanism of targeting histone deacetylase 1 (HDAC1) regulating cervical cancer cell migration and invasion. Methods Clinically collected 86 patients with cervical cancer. The levels of miR-513c-5p in tumor tissues and adjacent tissues were detected by Real-time PCR. The relationship between miR-513c-5p and pathological characteristics of cervical cancer was analyzed. It was verified that miR-513c-5p targets HDAC1 by a dual luciferase report. Cervical cancer HeLa cells were divided into four groups: control group, mimic group, mimic+HDAC1 group and HDAC1 group. MiR-513c-5p and(or) HDAC1 were overexpressed by plasmid transfection technology. Real-time PCR and Western blotting were used to detect the expression level of RNA or protein, respectively. The cell growth, migration, and invasion capabilities of each group were measured by CCK-8 method, cell scratch test, and Transwell test. Results The level of miR-513c-5p in cervical cancer tissues was significantly lower than that in adjacent tissues. Low levels of miR-513c-5p were associated with higher local invasion, lymphatic metastasis, and distal metastasis (P<0. 05). MiR-513-5p targeted HDAC1 expression. Overexpression of miR-513c-5p inhibited significantly the growth, migration and invasion of cervical cancer cells (P < 0. 05). Overexpression of HDAC1 promoted growth, migration and invasion (P<0. 05), and reversed the inhibitory effect of miR-513c-5p (P<0. 05). Conclusion Low levels of miR-513c-5p might be related to cervical cancer metastasis, and miR-513c-5p could inhibit the growth, migration and invasion of cervical cancer HeLa cells by targeted inhibition of HDAC1 protein expression.
ABSTRACT
<p><b>OBJECTIVE</b>The influence of processing methods on chemical constituents in Radix Paeoniae Alba was observed.</p><p><b>METHOD</b>A HPLC method was used for analyzing the changes of eight major constituents, namely gallic acid, paeoniflorin sulfonate, catechin, paeoniflorin sulfonate, albiflorin, paeoniflorin, benzoic acid, pentagalloylglucose and benzoylpaeoniflorin, with the three processing procedures of decorticating, boiling and fumigating by burning of sulphur. Analysis was performed using a Zorbax SB-C18 column (4.6 mm x 250 mm, 5 microm) with the mixture of acetonitrile (A) and 0.015% phosphoric acid solution as mobile phase in gradient mode. The detection wavelength was set at 230 nm and the column temperature was at 30 degrees C.</p><p><b>RESULT</b>Except for gallic acid and pentagalloylglucose, the other constituents decreased during procedure of decorticating and boiling. Fumigating by burning of sulphur would produce a new compound, paeoniflorin sulfonate, which was a byproduct from the reaction of paeoniflorin with SO2.</p><p><b>CONCLUSION</b>The significant changes were produced in chemical constituents of Radix Paeoniae Alba during three processing procedures. Therefore, the processing of Radix Paeoniae Alba should be strictly controlled and standardized.</p>
Subject(s)
Benzoates , Chemistry , Bridged-Ring Compounds , Chemistry , Chromatography, High Pressure Liquid , Gallic Acid , Glucosides , Chemistry , Hot Temperature , Hydrolyzable Tannins , Molecular Structure , Monoterpenes , Paeonia , Chemistry , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Sulfur , Technology, Pharmaceutical , MethodsABSTRACT
<p><b>AIM</b>To establish the method of HPLC-fingerprint analysis for the quality control of Dalbergia odorifera and identify its main constituents by HPLC-MS.</p><p><b>METHODS</b>The 37 hatches of samples were analyzed on a Phenomenex Luna C18 column with a gradient of acetonitrile and 0.3% aqueous acetic acid at a flow rate of 1.0 mL x min(-1) and detected at 275 nm. Furthermore, the typical samples were detected by HPLC-DAD-MS under negative ion mode.</p><p><b>RESULTS</b>37 batches of D. odorifera samples were classified into three types based on the results of similarity analysis. According to the comparison of the tR, MS data and UV maximum absorbance (gamma(max)) values with the standards, 10, 7 and 2 phenolic components were identified in three types of D. odorifera extracts, separately.</p><p><b>CONCLUSION</b>The method is repeatable and reliable, and it is capable of effectively controlling the quality of D. odorifera.</p>