ABSTRACT
Background: Scutellaria barbata D. Don (SB) is a widely used herbal medicine in China, with various pharmacological effects such as anti-oxidation, anti-inflammation, and anti-cancer. This work is aimed to investigate the tumor-suppressive effect of SB in cervical cancer (CC) and to identify its underlying mechanisms. Methods and materials: CC cell lines (HeLa and HT-3) were treated with different concentrations of SB chloroform extract (ECSB) (0, 0.2, 0.5 mg/ml). MiR-195-5p and LOXL2 mRNA expression in CC cell lines and tissue samples was detected by quantitative real-time polymerase chain reaction. Cell counting kit-8 experiment was utilized to examine cell viability; TUNEL assay and Transwell experiment was executed to examine cell apoptosis, migration, and invasion. Western blotting experiments were implemented to detect LOXL2 protein expression. Bioinformatics and dual-luciferase reporter gene experiment were executed to examine the binding relationship between miR-195-5p and LOXL2. Results: ECSB repressed the viability, migration, and invasion of HeLa and HT-3 cells, and promoted cell apoptosis in a dose-dependent manner. MiR-195-5p was remarkably under-expressed in CC tissues and cells, and ECSB up-regulated miR-195-5p expression. MiR-195-5p inhibitors partially counteracted the suppressive effects of ECSB on the malignant phenotypes of HeLa and HT-3 cells. LOXL2 was a downstream target of miR-195-5p, and ECSB up-modulated LOXL2 expression by specifically repressing miR-195-5p. Conclusion: SB restrains CC cell proliferation and metastasis and promotes cell apoptosis via miR-195-5p/LOXL2, which may be a potential therapeutic agent for CC treatment.
ABSTRACT
According to the specific construction of the pSUPER. retro. puro vector, RNA interfering with the hypoxia-inducible factor 1alpha (HIF-1alpha) plasmid was constructed in this research programme. The efficiency was (87.15 +/- 2.36)% and green fluorescent protein was observed after 36 hours when the constructed plasmid co-transformed into the prostatic carcinoma cell line PC-3; compared with the control groups, this constructed plasmid could reduce the expression of total RNA and protein in PC-3 cells significantly after 48 hours. The cells were checked out by the plasmid resistance against the puromycin biotin, the clones of which were selected and enlarged for two weeks, then the cell strain of pSUPER-siHIF-1alpha/PC-3 was collected. The HIF-1alpha protein expression of the pSUPER-siHIF-1alpha/ PC-3 cells of also decreased significantly. The results showed that the RNA interference could be used in the construction of expression vector of constructed siRNA inhibiting the expression of HIF-1alpha with PC-3 cells, which is the basis of researching the pathology, multiplication, and metastasis of HIF-1alpha in the prostatic carcinoma and other cancers.
