Study On The Effect Of Mulberry Active Substance Moracin D In Inhibiting Proliferation And Inducing Apoptosis In Gastric Cancer

The Gastric cancer’s incidence and mortality are at the head of malignant tumors.The prognosis of gastric cancer remains poor despite the fact that surgery,chemotherapy and radiotherapy have extended survival time.Therefore,new strategies are needed.The treasure trove of Chinese herbal medicine,with long history and abundant resources,has been acknowledged as a source of research and development for new drugs.Additionally,the natural products of Chinese herbal medicines have become a highly sought-after subject in anti-cancer studies.Moracin D is a benzofuran compound derived from the mulberry tree.Studies have proved that Moracin D has anti-inflammatory,antiviral and metabolic improvement pharmacological effects.Moracin D’s function in prostate cancer and breast cancer has been established.However,the role of Moracin D in gastric cancer is not yet known.Therefore,this article will delve into the role of Moracin D in gastric cancer and its mechanism.Here are the main results.1.Moracin D inhibits the proliferation of gastric cancer cellsBGC-823,HGC-27,SGC-7901,MKN-45 and GES-1 were selected for preliminary exploration experiments.The IC₅₀ values were measured at 25.37μM,15.01μM,19.51μM,20.44μM,and 111.6μM,respectively.Therefore,three cell lines with the highest,lowest and medium sensitivity to Moracin D were selected for experiment,namely HGC-27,BGC-823 and MKN-45,and three drug concentration gradients of 10μM,20μM and 40μM were set.The control group was DMSO with a concentration of 40μM.MTT experiment,Brd U experiment,and plate cloning experiment all showed that with the increase of dosing concentration,the proliferation of gastric cancer cells was weakened.2.Moracin D inhibits the tumorigenesisi ability of gastric cancerIn vitro,Moracin D can inhibit the proliferation of gastric cancer cells.Does Moracin D affect the growth of gastric cancer in vivo?We first performed a soft agar experiment to simulate a semi-solid environment in vivo.The results demonstrated that,in comparison to the control group,monoclonals’number in the experimental group was reduced.The clone size was also smaller than the control group.We then further established an animal subcutaneous transplant tumor model.Gastric cancr cells were injected into nude mice to form progressive tumors to detect the tumorgenesis ability of cells.The results revealed that the tumors in Moracin D group were significantly smaller in terms of tumor volume and tumor weight.Ki67is seen clinically as a marker of the degree of cellular malignancy.The expression level of ki67was also tested.The ki67 positivity rate was low in the Moracin D group.This shows the tumorigenesis inhibition of Moracin D.3.Moracin D blocks the gastric cancer cell cycle in the G2/M phaseCell cycle was analyzed by Flow cytometry,and Moracin D treatment caused a blockage of the G2/M phase.Western bolt experiments were then conducted to measure the expression of G2/M-related proteins,and the results indicated a decrease in CDK2 and cyclin B1 protein levels.Levels of these proteins varied in concentration-dependent and time-dependent.Moracin D may regulate the cell cycle by proteins in relation to cycle.4.Moracin D induces apoptosis of gastric cancer cellsInducing apoptosis is a major strategy for the treatment of cancer.In the experiment,we found that the cells treated with Moracin D have the characteristics of apoptosis.Therefore,we quantified apoptosis using flow cytometry.A marked increase in apoptosis rate of gastric cancer cells was observed in the Moracin D treated group when compared with the control group.Furthermore,C-PARP,C-caspase3 and C-caspase7 protein levels were raised following Moracin D treatment.Bcl-2,a protein that is essential for cancer suppression in traditional Chinese herbal medicines,was significantly reduced.Therefore,we suspect that Bcl-2 may be a target for Moracin D to inhibit gastric cancer cell proliferation and induce apoptosis.5.Overexpression of Bcl-2 can save the effect of Moracin D on gastric cancer cellsTo verify this hypothesis,we used lentivirus infection assays to overexpress Bcl-2 in three gastric cancer cell lines.MTT,Brd U,and plate cloning experiments showed that the proliferation inhibition induced by Moracin D was partially reversed.Overexpression of Bcl-2partially restored the proliferative capacity of gastric cancer cells.At the same time,Western blot experiments also found that the expression of C-PARP,C-caspase3,C-caspase7 proteins was also partially restored.Overexpression of Bcl-2 partially reverses the effect of Moracin D on apoptosis-associated proteins.6.Moracin D combined with 5-fluorouracil（5-FU）inhibits the growth of gastric cancer cellsMoracin D can inhibit the proliferation of gastric cancer cells,so what role will it play when combined with 5-FU,a common clinical treatment for gastric cancer?Therefore,we conducted a combination drug experiment.Experiments utilizing MTT revealed that the combination of Moracin D and 5-FU inhibited cell proliferation.The Jin’s formula analysis proved that the inhibition is synergistic.Brd U also demonstrated the synergistic inhibition of Moracin D with 5-FU.The Western blot experiment found that the apoptosis effect caused by the combination of the two drugs was more obvious.In summary,Moracin D shows anticancer activity both in vivo and in vitro.Moracin D can inhibit gastric cancer proliferation and induce apoptosis.Theoretically,this provides a basis for clinical use in treating gastric cancer.