Study On Petroleum Ether Extract Of Scutellaria Barbata D. Don And Hedyotis Diffusa Willd Joint DACH1 Anti-endometrial Cancer Activity And Related Mechanism

Objective: To investigate the chemical components and the activity of anti-endometrial cancer of the extract in Scutellaria barbata D. Don and Hedyotis diffusa Willd and the herb pair. And through chemical composition analysis and contrast of the active site, initially speculated that the correlation of Scutellaria barbata D. Don, Hedyotis diffusa Willd anti-endometrial cancer activity and chemical composition.To study the anti-endometrial cancer activity and mechanism of Hedyotis diffusa Willd active site, DACH1 and two joint. Initially revealed the possible molecular mechanisms of inhibition of endometrial carcinoma cell proliferation and induced apoptosis by Hedyotis diffusa Willd active site and DACH1,and provide an important basis for the molecular diagnosis and treatment of endometrial cancer.Method: Volatile oil and petroleum ether extract in Scutellaria barbata D. Don and Hedyotis diffusa Willd and the herb pair were determined by the GC-MS technology. Activity of anti-endometrial cancer cells was researched by MTT assay, and screening out the active site. The recombinant lentivirus LV-DACH1-GFP which DACH1 overexpression was infected into human endometrial carcinoma HEC-1A cells. Further, the rate of clone formation, cell migration, cell cycle and the expression of FASN were detected, and the inhibitory effect of DACH1 on endometrial cancer was detected by tumor formation in nude mice. Petroleum ether extract of Hedyotis diffusa Willd(PEEHDW)used in HEC-1A cells. The clone forming ability of cells was detected by plate clone experiment and scratches and cell migration test cell migration ability. The cell cycle and Casepase-3 expression was measured by flow cytometry. The expression of Cyclin D1、Bcl-2、FASN was examined by western blot. The proliferation of Endometrial Cancer Cells was evaluated in tumor formation in nude mice and the pathomorphological changes of tumor was observed by H-E staining. The combined of PEEHDW and DACH1 in HEC-1A/ DACH1 cells, Further, the rate of clone formation, cell migration, tumor formation in nude mice were detected.Results: The minimum value of the IC50 for HEC-1A cells of Volatile oil and petroleum ether extract in Scutellaria barbata D. Don and Hedyotis diffusa Willd and the herb pair was 38.532 μg·m L-1. Unsaturated fatty acid, esters(26.926%),anthraquinones(14.762%), sterols(10.286%),flavone(1.410%),terpenes and lactones(0.206%),and other compounds. After recombinant lentivirus LV-DACH1-GFP, DACH1 could be obtained mainly in nucleus. The expression levels of DACH1 m RNA and protein of HEC-1A cells in LV-DACH1-GFP infection group were higher than those in control group and blank group. The proliferation of HEC-1A cells in LV-DACH1-GFP infection group was inhibited by MTT. The experimental results of the rate of clone formation and cell migration showed that the ability of cell clone formation and cell migration was inhibited significantly,and the cell cycle was arrested at the G0/G1 phase. Western blot results showed that the high expression of DACH1 inhibited FASN expression. The results of tumor formation in nude mice showed that DACH1 overexpression inhibited the in vivo proliferation of HEC-1A cells to a certain extent. PEEHDW inhibited the proliferation of HEC-1A cells and clone formation and migration of ability as medicine concentration increased. Petroleum ether extract blocked the cell cycle of HEC-1A at the G0/G1 phase and induced cell apoptosis by promoting the expression of Caspase-3. Western blot was used to detect the PEEHDW inhibit FASN, Cyclin D1 and Bcl-2 protein expression, promote the expression of DACH1 protein, And the PEEHDW on HEC-1A tumor growth showed significant inhibition effect. The PEEHDW was used in HEC-1A/DACH1 cells, which can significantly inhibit the proliferation ofr HEC-1A/DACH1 cells in vitro and in vivo, clone formation and migration ability, and promote the Caspase-3 expression to induce cell apoptosis.Conclusion: The PEEHDW has the strongest activity of anti-endometrial cancer through MTT assay. It was speculated that the higher content of anthraquinones could be related to the strongest activity. DACH1 gene may inhibit HEC-1A cells clone formation and migration ability, block HEC-1A cell growth cycle, inhibit the expression of FASN,regulate the production of lipid signaling, resulting in suppression of proliferation of endometrial cancer cells in vitro and in vivo.PEEHDW inhibit cell proliferation of endometrial cancer cells HEC-1A by inhibiting Cyclin D1 protein expression, probably by reducing the Bcl-2 expression and activation of Caspase-3 to induce cell apoptosis, may also inhibit FASN expression inhibition of endometrial cancer cell growth and induce apoptosis of HEC-1A. The PEEHDW and DACH1 combined with anti endometrial cancer activity is more obvious.