1.Constructing an expression vector for human lncRNA H19 and the effect of its overexpression on MCF-7 cell proliferation
Yan PENG ; Haitang XIE ; Hong SUN ; Ying ZENG ; Qiongni ZHU ; Tailin LI ; Guo WANG ; Yuanshan ZHU
Chinese Pharmacological Bulletin 2015;(4):555-559,560
Aims To construct an expression vector of human lncRNA H 1 9 ,and to determine the effect of H1 9 overexpression on MCF-7 cell proliferation. Methods Total RNA was extracted from MCF-7 cells,and the full-length of H1 9 lncRNA was amplified by RT-PCR and subcloned into pcDNA3.1 (-)ex-pression vector.The constructed H1 9 expression vector was transfected into HEK-293T and COS-7 cells and the H1 9 lncRNA expression was evaluated by real-time PCR.Following the transfection of H1 9 expression vec-tor into MCF-7 cells for 0,24h and 48h and H1 9 siR-NA interference fragment into MCF-7 cells for 24h, MCF-7 cell proliferation was determined by MTS as-say.Results A hH1 9-pcDNA3.1 (-)expression vector was successfully constructed. At Forty-eight hours after the transfection with H1 9 expression vector in to MCF-7 cells,cell proliferation was significantly increased in the transfected group compared to those without transfection and to those transfected with a neg-ative control vector,while twenty-four hours after the transfection with H1 9 siRNA interference fragment into MCF-7 cells,cell proliferation was significantly de-creased in the transfected group compared to those transfected with a negative control vector.Conclusion Ectopic overexpression of H1 9 lncRNA can promote breast cancer MCF-7 cell proliferation.
2.Study on Different Components of Rhei Radix Et Rhizoma from 3 Different Origins Based on HPLC Fingerprint and UPLC- Q-TOF-MS
Dandan LI ; Hui ZHANG ; Siyu LI ; Tailin GUO
China Pharmacy 2019;30(23):3240-3245
OBJECTIVE: To establish HPLC fingerprints of Rhei Radix Et Rhizoma from different origins, and identify the differential components qualitatively. METHODS: HPLC fingerprints of 30 batches of Rhei Radix Et Rhizoma (20 batches of Rheum palmatum, 5 batches of Rheum tanguticum and 5 batches of Rheum officinale) were established and similarity evaluation was performed by using Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012 edition). Qualitative identification of differential components of Rhei Radix Et Rhizoma from 3 different origins were performed by using PLS-DA combined with UPLC-Q-TOF-MS. RESULTS:The fingerprint similarities of 30 batches of samples were between 0.609 and 0.960. According to PLS-DA analysis, Rhei Radix Et Rhizoma were significantly aggregated into 3 groups according to the origin. There were 18 different components among 3 groups, which were identified by UPLC-Q-TOF-MS as resveratrol-4′-O-β-D-(6″-O-gallacyl)-glucoside, lindleyin, rhein-8-O-glucoside, epicatechin gallate, 4-(4′-hydroxyphenyl)-2-butanone-4′-O-β-D-(2″-O-cinnamyl-6″-O-gallacyl)-glucoside. CONCLUSIONS: Established method can effectively identify Rhei Radix Et Rhizoma from 3 different origins, and the differential components can be distinguished, which provides a reference for the identification and quality evaluation of multi-source medicinal materials.