Research progress on the anti-tumor effect of oridonin / 国际中医中药杂志

Oridonin is one of the main anti-tumor active ingredients of Rabdosiae Rubescentis Herba. In the development process of a variety of malignant tumors, such as liver cancer, breast cancer, cervical cancer, pancreatic cancer, lung cancer, gastric cancer, esophageal cancer, colon cancer, etc, it has a significant inhibitory effect. Oridonin plays the rold of anti-tumor effect mainly by inhibiting tumor cell proliferation, inducing tumor cell apoptosis, inhibiting tumor cell invasion and migration, inducing tumor cell autophagy, reducing telomerase activity and reversing the drug resistance of the tumor cell.

Study on the mechanism of oridonin to reverse drug resistance of breast cancer cell MCF- 7 to fulvestrant / 中国药房

OBJECTIVE To explore the mechanism of oridon in（Ori）reversing the drug resistance of breast cancer cell MCF-7 to fulvestrant （Ful）. METHODS Ful-resistant breast cancer cell strains MCF- 7/Ful were induced and constructed in vitro . The relative cell viability of MCF- 7 cells and MCF- 7/Ful cells was detected by MTT assay ，inhibitory rate of Ori to MCF- 7/Ful cells was also detected. CompuSyn software was used to analyze the synergistic effect of Ori and Ful. MCF- 7/Ful cells were randomly divided into blank control group ，Ful group （5 μmol/L），Ori group （8 μmol/L），Ful（5 μmol/L）+Ori（8 μmol/L）group. The phosphorylation of phosphatidylinositol 3 kinase（PI3K）/protein kinase B （Akt）signaling pathway related protein in each group was detected by Western blot. MCF-7/Ful cells were used to prepare drug resistance model of transplanted tumor in nude mice，and they were randomly divided into blank control group ，Ful group （80 μmol/g），Ori group （50 μmol/g），Ful（80 μmol/g）+ Ori（50 μmol/g）group. The tumor weight and tumor inhibition rate were calculated ，to verify the reversal effect of Ori and Ful in vivo. RESULTS MTT assay showed that when Ful ≥10 μmol/L，the relative cell viability of MCF- 7/Ful cells was significantly higher than that of MCF- 7 cells（P＜0.05），and the drug resistance was significantly enhanced ；Ori had a significant inhibitory effect on MCF- 7/Ful cells ，the inhibition rate of Ori combined with Ful to MCF- 7/Ful cells was significantly increased （P＜0.05 or P＜0.01），and the effect of reversing drug resistance was significantly increased . The results of Western blot showed that compared with Ful group ，the phosphorylation levels of PI 3K and Akt protein in Ful+Ori group were significantly decreased （P＜0.05 or P＜ 0.01）. In vivo results showed that the tumor volume and mass of Ful+Ori group were significantly decreased ，and the tumor inhibition rate was （63.90±4.11）％，which was significantly higher than that of Ful group （P＜0.01）. CONCLUSIONS Ori can reverse drug resistance of breast cancer cell MCF- 7 to Ful ， and this effect may be through regulating PI 3K/Akt signaling pathway.

Effects of Oridonin-loaded Chitosan Graphene Oxide on the Proliferation and Apoptosis of A 549 Cells / 中国药房

OBJECTIVE：To study the effects of chitosan graphene oxide car rier（CS-GO）loaded with oridonin （CS-GO- oridonin）on the proliferation and apoptosis of human lung cancer A 549 cells. METHODS ：Taking A 549 cells as objects ，the survival rate of cells were detected by CCK- 8 method after treated with different concentrations of CS-GO （3，6，12，24，48 μg/mL）and CS-GO-oridonin loaded with same mass of oridonin （3，6，12，24，48 μg/mL，by the weight of oridonin ，the same below）. IC 50 of CS-GO-oridonin was calculated. The cell morphology were observed by microscope after treated with CS-GO and CS-GO-oridonin（both 32 μg/mL）for 2，4，10，24 h. The uptake of CS-GO ，oridonin，CS-GO-orionin（all 32 μg/mL）by cells was observed with fluorescence labeling method. The apoptosis of cells and the content of ROS were observed by flow cytometry after treated with different concentrations of CS-GO （16，32，64 μg/mL）and CS-GO-oridonin （16，32，64 μg/mL）. The expression of anti-apoptosis related proteins （Mcl-1，Bax and Bak ）were detected by Western blot. RESULTS ：After treated with different concentrations of CS-GO ，the survival rate of cells was still above 90％ ；after treated with different concentrations of CS-GO-oridonin，the survival rate of cells showed a downward trend ，and was significantly lower than that of CS-GO group （P＜ 0.01）；IC50 of CS-GO-oridonin was 32.61 μg/mL. After CS-GO treatment，the cell morphology did not change significantly ；after CS-GO-oridonin treatment ，the cells shrunk and fell off in clusters ，and the suspended matter increased ；the fluorescence of oridonin and CS-GO-orionin taken up by cells was enhanced than CS-GO. Compared with blank group ，there was no significant change in the apoptosis rate ，the content of ROS and the expression of apoptosis-related protein in 16，32，64 μg/mL CS-GO groups（P＞0.05）；apoptosis rate ，the content of ROS ，the protein expression of Bax and Bak in 16，32，64 μg/mL CS- GO-oridonin groups were increased significantly ，while the protein expression of Mcl- 1 were decreased significantly. Above indexes were significantly high er or lower than the same concentration CS-GO group （P＜0.05）. CONCL USIONS：CS-GO dose not affect the proliferation and apoptosis of A 549 cells；CS-GO-oridonin has obvious inhibition and apoptosis promoting effect on cells ，which may be related to increasing ROS production and regulating the expression of apoptosis related proteins.

Inhibitory effect of oridonin on proliferation and migration of human esophageal squamous cancer cell lines KYSE-150 and KYSE-450 / 解剖学报

Objective To explore the effect of oridonin (ORI) on proliferation, apoptosis, cell cycle and migration of esophageal squamous carcinoma cell (ESCC) lines KYSE-150 and KYSE-450. Methods The effect of ORI on the proliferation and clony formation of esophageal cancer cells were detected by MTT and colony formation assays. Flow cytometry was performed to examine the impact of ORI on cell apoptosis and cell cycle. Transwell assay was applied to detect the role of ORI on cell migration. The effect of ORI on the expression of anti-apoptotic protein Bcl-2, cell cycle inhibitory protein p21Cip1/Waf1, epithelial-mesenchymal transition (EMT) related markers were examined by Western blotting. Results ORI had a significant inhibitory effect on the proliferation, migration and clone formation of KYSE-150 and KYSE-450 cells (P<0. 05) in a time and dose-dependent manner. With the increase of ORI concentration, apoptosis rate and the proportion of cells in G2/M phase increased significantly (P<0. 05), and the proportion of cells in G0/G1 phase decreased significantly (P < 0. 0 5). Bcl-2, vimentin and p-catenin were down-regulated and p21Cipl/Wafl, Ecadherin were up-regulated after treatment of ORI on ESCC cells for 48 hours. Conclusion ORI may inhibit ESCC cell proliferation and clony formation by inducing apoptosis and resting cells in G2/M phase, and suppress ESCC cell migration via inhibiting EMT process.

Preparation Technology Study on Oridonin A Oral Liposomes Based on Supercritical Fluid Solution-enhanced Dispersion / 中国药房

OBJECTIVE： To optimize the preparation technology of Oridonin A oral liposomes （ORI-LIP） by using supercritical fluidsolution-enhanced dispersion （SEDS） technology， and to investigate its advantage with routine liposome preparation technologies. METHODS： Using particle size as evaluation index， orthogonal design was employed to investigate the influence of pressure， temperature and flow rate on the preparation technology of ORI-LIP by SEDS. At the same time， thin film dispersion and reverse evaporation method were used to prepare ORI liposomes. The particle size， encapsulation efficiency， drug loading amount and stability （accelerated test for 6 months） were compared among 3 methods. Moreover， the difference in dissolution behavior in vitro of ORI crude drug and 3 kinds of liposomes was evaluated. RESULTS： The optimized preparation condition of ORI liposomes by SEDS included temperature of 50 ℃， pressure of 18 MPa， flow rate of 1 mL/min. Compared with thin film dispersion and reverse evaporation method， the liposomes prepared by the SEDS method exhibited smaller particle size [（147.4±4.8）nm]， better encapsulation efficiency （67.8％）， drug-loading amount （7.8％） and stability （particle size increased slightly， encapsulation efficiency decreased only by 4.4％）. Results of in vitro dissolution test showed that compared with crude drug， release rate of each liposome was slow and persistent， and the cumulative release rate was higher. The accumulative release rate of ORI-LIP prepared by SEDS could achieve to 67.2％， and reached to dissolution equilibrium at 24 h. CONCLUSIONS： ORI-LIP prepared by SEDS has smaller particle size， higher encapsulation efficiency， drug loading amount and stability， which can improve the in vitro release of ORI. Compared with conventional methods， SEDS technology has certain advantages.

Homoharringtonine synergy with oridonin in treatment of t(8; 21) acute myeloid leukemia / 医学前沿

Collaboration of c-KIT mutations with AML1-ETO (AE) has been demonstrated to induce t(8; 21) acute myeloid leukemia (AML). Targeted therapies designed to eliminate AE and c-KIT oncoproteins may facilitate effective treatment of t(8; 21) AML. Homoharringtonine (HHT) features activity against tumor cells harboring c-KIT mutations, whereas oridonin can induce t(8; 21) AML cell apoptosis and AE cleavage. Therefore, studies should explore the efficacy of combination therapy with oridonin and HHT in t(8; 21) AML. In this study, we investigated the synergistic effects and mechanism of oridonin combined with HHT in t(8; 21) AML cell line and mouse model. The two drugs synergistically inhibited cell viability and induced significant mitochondrial membrane potential loss and apoptosis. Oridonin and HHT induced significant downregulation of c-KIT and its downstream signaling pathways and promoted AE cleavage. HHT increased intracellular oridonin concentration by modulating the expressions of MRP1 and MDR1, thus enhancing the effects of oridonin. The combination of oridonin and HHT prolonged t(8; 21) leukemia mouse survival. In conclusion, oridonin and HHTexert synergistic effects against t(8; 21) leukemia in vivo and in vitro, thereby indicating that their combination may be an effective therapy for t(8; 21) leukemia.

Oridonin induces growth inhibition and apoptosis in human gastric carcinoma cells by enhancement of p53 expression and function

The tumor suppressive role of oridonin, an active compound extracted from Rabdosia rubescens, has been proven in several gastric cancer (GC) cell lines. The present study aimed to evaluate the effect of oridonin on another GC cell line, SNU-216, and explore the potential mechanisms. The viable cell numbers, cell migration, survival fraction, and cell viability were, respectively, evaluated by trypan blue exclusion assay, wound healing assay, clonogenic assay, and CCK-8 assay. Cell apoptosis was determined by flow cytometry assay and western blot. The expression of p53 was inhibited by transient transfection, and the efficiency was verified by western blot. qRT-PCR was performed to measure the mRNA expression of p53. Western blot was used to evaluate the protein expression of apoptosis, DNA damage and p53 function related factors. We found that oridonin significantly inhibited cell proliferation, migration, and survivability, and enhanced cell apoptosis in SNU-216 cells. However, it had no influence on HEK293 cell viability. Oridonin also remarkably enhanced the anti-tumor effect of cisplatin on SNU-216 cells, as it significantly increased apoptotic cells and decreased cell viability. Moreover, the mRNA and protein expression of p53 was significantly up-regulated in oridonin-treated cells, while Mdm2 expression was down-regulated. Furthermore, oridonin enhanced p53 function and induced DNA damage. Knockdown of p53 or employing the caspase inhibitor, Boc-D-FMK, reversed the effect of oridonin on cell viability and apoptosis-related protein expression. The present study demonstrated that oridonin exhibited an anti-tumor effect on GC SNU-216 cells through regulating p53 expression and function.

The Preparation and In Vitro Studies of Phytantriol-Based Cubosomes Containing Oridonin / 中国药学杂志

OBJECTIVE: To prepare oridonin-loaded cubosomes, and encapsulate oridonin(ORI) for the purpose of enhancing the solubility and prolonging the action time. METHODS: Phytantriol (PYT) was firstly used to cooperate with Poloxamer 407-propylene glycol-water system to improve the solubility of ORI for developing ORI-loaded cubosomes. Polarizing microscope, small angel X-ray scattering (SAXS) and cryogenic transmission electron microscopy(cryo-TEM) were used to study the characters of cubosomes. RESULTS: Under homogenization conditions of 1 200 bar for 9 cycles, the obtained PYT-based cubosomes had narrow particle size distribution with a mean particle size of (225.9 ± 5.6) nm. The internal structures of cubosomes were revealed by small-angle X-ray scattering as a bicontinuous cubic liquid crystalline phase with Pn3m geometry. The encapsulation efficiency and drug loading determined by ultrafiltration centrifugation were (86.6 ± 1.5)% and (3.69 ± 0.06) mg•g-1, the solubility of ORI had been increased by 5.20 times. CONCLUSION: The optimized formulas of cubosomes show obvious 24 h-sustained release, and the in vitro release profiles fitt the Higuchi release model well, implying diffusion-control as main release mechanism.

Effects and mechanism of oridonin on invasion and migration abilities of human melanoma A375 cells / 中草药

Objective To study the effects and the mechanism of oridonin on inhibiting the invasion and migration abilities of human melanoma A375 cells. Methods Human melanoma A375 cells were cultured and treated respectively with indicated concentrations of oridonin by cell culture technique. The proliferation rate was detected by CCK-8 method. The migration ability was measured by wound healing assay. The invasion ability was examined by Transwell assay. The adhesion capabilities were evaluated by adhesion assay. The epithelial-mesenchymal transition (EMT) and matrix metalloproteinases (MMPs) related protein expression levels were determined by Western blotting. Results CCK-8 assay showed the median inhibition concentration (IC50) of 48 h was 47.94 μmol/L. Oridonin (5, 10, and 20 μmol/L) inhibited the migration, invasion and adhesion abilities of human melanoma A375 cells in a dose-dependent manner (P < 0.05). After oridonin treatment, the protein expression levels of E-cadherin increased significantly (P < 0.05) and the protein levels of Snail, N-cadherin, vimentin, MMP-2, and MMP-9 decreased significantly (P < 0.05). Conclusion Oridonin inhibits the migration, invasion and adhesion abilities of human melanoma A375 cells. The mechanism may be related with the regulating effects of oridonin on EMT and MMPs.

Protective effects of oridonin on adriamycin-induced myocardial apoptosis through inhibiting autophagy / 局解手术学杂志

Objective To investigate the effects and mechanisms of oridonin on adrimycin-induced myocardial apoptosis. Methods Cells were divided into H9C2,adriamycin,oridonin(5 μM),oridonin(10 μM) and oridonin(20 μM) group.Cells were treated with adriamy-cin except H9C2 group and cells in oridonin(5,10,20 μM)group were treated with oridonin at the same time.The concentrations of superox-ide dismutase(SOD) and malondialdehyde(MDA) were detected and the cell proliferation was detected by CCK8 assay.Cell apoptosis was determined by flow cytometry.The expressions of autophagy-related proteins(Beclin1,P62 and LC3) were measured by western blot.And the expression of LC3 also detected by immunofluoresence.Results Compared with H9C2 group,the concentration of SOD decreased and MDA increased in adriamycin group;compared with adriamycin group,SOD increased but MDA decreased in oridonin(5,10,20 μM)group signifi-cantly.Meanwhile,cell proliferation was inhibited and apoptosis was induced in adriamycin group compared with H9C2 group;but cell prolif-eration rate was increased and apoptosis rate was decreased in oridonin(5,10,20 μM)group compared with adriamycin group.In addition,ad-riamycin up-regulated the protein level of Beclin1 and ratio of LC3Ⅱ/LC3Ⅰ,and inhibited the expression of P62;oridonin(5,10,20 μM) attenuated the effects of adriamycin on Beclin1,ratio of LC3Ⅱ/LC3Ⅰand P62 notably.Conclusion Oridonin alleviated adriamycin-indued myocardial apoptosis by inhibiting autophagy.

Mechanism of Oridonin-induced Apoptosis of Glioma SHG44 Cells / 中国医科大学学报

Objective To study the molecular mechanism of oridonin-induced apoptosis of ghoma SHG44 cells.Methods A growth curve was plotted using CCK-8 colorimetric method with different concentrations of oridonin (0,1.25,2.5,5,10,20,and 40 μmol/L)to observe its effect on the growth of SHG44 cells.Hoechst33258 and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were used to examine the changes in cell morphology and flow cytometry was used to detect cell apoptosis.Western blotting was used to analyze the expression of apoptosis-related proteins (Caspase-3,cleaved Caspase-3,Bax,and Bcl-2)in SHG44 cells.Results SHG44 cell proliferation was significantly suppressed after 24 and 48 h Oridonin treatment,with a half-maximal inhibitory concentration of 7.865 and 4.74 μmol/L,respectively.Hoechst33258 and TUNEL staining showed changes in cell morphology such as shrinkage and nucleus fragmentation and morphogenesis,which are indicative of apoptosis.Western blotting analysis showed that oridonin inhibited the expression of Bcl-2 and activated the expression of Caspase-3 and Bax.Conclusion Oridonin can inhibit the proliferation and induce the apoptosis of SHG44 cells by regulating the expression of apoptosis-related proteins.

Preparation of Oridonin-Single-Walled Carbon Nanotubes and Study on Its Adsorption Kinetics / 医药导报

Objective To investigate the method for preparing oridonin-single-walled carbon nanotubes ( ORI-SWCNTs) nanocomposite and study its adsorption kinetics. Methods ORI-SWCNTs nanocomposite was prepared by using the method of solution mixing. The synthesized ORI-SWCNTs nanocomposite was characterized by using Laser particle size analyzer, Fourier transform infrared, DSC analysis, powder X-ray diffraction and electron microscopy techniques. Results The encapsulation efficiency and loading capacity of ORI in SWCNTs-COOH nanocarrier was estimated to be about (70.23±2.1) %and (27.29±1.2) %, respectively. The Zeta potential was (-34.29±1.2) mV, partical size was about (458±18) nm. The absorption of ORI on SWCNTs-COOH could be explained by pseudo-second-order model. Conclusion The established preparation process of ORI-SWCNTs nanocomposite by solution mixing is feasible, with higher loading efficiency and encapsulate efficiency..

Preparation of Oridonin-Single-Walled Carbon Nanotubes and Study on Its Adsorption Kinetics / 医药导报

Objective To investigate the method for preparing oridonin-single-walled carbon nanotubes ( ORI-SWCNTs) nanocomposite and study its adsorption kinetics. Methods ORI-SWCNTs nanocomposite was prepared by using the method of solution mixing. The synthesized ORI-SWCNTs nanocomposite was characterized by using Laser particle size analyzer, Fourier transform infrared, DSC analysis, powder X-ray diffraction and electron microscopy techniques. Results The encapsulation efficiency and loading capacity of ORI in SWCNTs-COOH nanocarrier was estimated to be about (70.23±2.1) %and (27.29±1.2) %, respectively. The Zeta potential was (-34.29±1.2) mV, partical size was about (458±18) nm. The absorption of ORI on SWCNTs-COOH could be explained by pseudo-second-order model. Conclusion The established preparation process of ORI-SWCNTs nanocomposite by solution mixing is feasible, with higher loading efficiency and encapsulate efficiency..

Effects of oridonin on invasion and migration of human hepatocellular carcinoma MHCC-97H cells / 中国病理生理杂志

AIM: To investigate the effects of oridonin on the invasion and migration of hepatocelluar carcinoma cells.METHODS: Human hepatocelluar carcinoma MHCC-97H cells were cultured and treated with 5, 10 or 20 μmol/L oridonin.The migration ability was measured by wound healing assay.The invasion ability was examined by Transwell invasion assay.The adhesion capabilities were evaluated by adhesion assay.The protein levels of LIM kinase-1 (LIMK-1), cofilin and phosphorylated cofilin (p-cofilin) were determined by Western blot.RESULTS: Oridonin inhibited the migration, invasion and adhesion abilities of MHCC-97H cells in a dose-dependent manner (P<0.05).After oridonin treatment, the expression of cofilin had no obvious change, but the protein levels of LIMK-1 and p-cofilin decreased significantly.CONCLUSION: Oridonin inhibits the invasion and migration of MHCC-97H cells.The mechanism may be related with the regulatory effect of oridonin on LIMK-1/cofilin signal transduction pathway.

Selection and identification of oridonin high yield strain and its antitumor activity / 中草药

Objective: To select and identify the endophytic fungi of Rabdosia rubescens as well as study on its antitumor activity in vitro in order to obtain the high yield strains of oridonin. Methods: Using the methods of TLC and HPLC, the oridonin high yield strain was selected from 256 strains of endophytic fungi which were isolated from the previous experiments. Oridonin of fermentation broth was determined by HPLC-MS. The strain was identified by morphological characteristics and ITS sequence methods and its antitumor activity in vitro was detected by MTT assay. Results: An oridonin high yield strain M-J-5 was obtained and identified as Penicillium oxalicum, which can inhibit the activity of human breast cancer cell line MCF-7. Conclusion: a oridonin high yield strain is obtained with antitumor activity and provides scientific basis for microbial production of oridonin and the development of new anticancer drugs.

Effect of oridonin on apoptosis and intracellular reactive oxygen species level in triple-negative breast cancer MDA-MB-231 cells / 中国中药杂志

Oridonin, which is an ent-kaurene diterpenoid isolated from traditional Chinese medicine Rabdosia rubescens, displays various bioactivities, including anti-inflammation, anti-bacteria and anti-tumor. This study aimed to investigate the effect of oridonin on apoptosis of triple-negative breast cancer MDA-MB-231 cells and its underlying mechanisms. The inhibitory effect of oridonin on proliferation of MDA-MB-231 cells was measured by MTT assay; Apoptosis was analyzed by flow cytometry with PI staining and Annexin V-FITC/PI staining; Intracellular reactive oxygen species (ROS) level was determined by ROS detection kit, and expressions of PARP, Bcl-2, caspase-3 were analyzed by Western blot. The results showed that oridonin exhibited a significant effect in inducing apoptosis of MDA-MB-231 cells, enhancing intracellular ROS level, down-regulating expression of Bcl-2 protein, and promoting cleavage of caspase-3 and its substrate PARP. These results indicated that the apoptosis-inducing effect of oridonin on MDA-MB-231 cells might be correlated with increase of intracellular ROS level, down-regulation of Bcl-2 protein and activation of caspase-3.

Therapeutic effect of oridonin on mice with prostate cancer

Objective: To investigate the therapeutic effect and the related mechanism of oridonin on mice with prostate cancer. Methods: Sixty BALB/C male nude mice were selected. A model of RM-1 cell transplantation tumor of prostate cancer was built by the subcutaneous inoculation of RM-1 cells. After that, those 60 experimental mice were randomly divided into groups A, B and C. Each group had 20 mice. Mice in group A were treated with 0.2 mL of normal saline (0.9%) by intraperitoneal injection once a day; mice in group B received intraperitoneal injection of 1.875 mg/mL of oridonin once a day; and mice in group C received intraperitoneal injection of 7.5 mg/mL of oridonin once a day. Mice in the three groups were treated uninterruptedly for 5 weeks and were all killed. Then, tumors were excised and weighed to calculate their growth inhibitory rate, volume increment and anti-tumor rate. Thymus and spleen of mice in the three groups were collected to calculate the thymus and spleen index. Immunohistochemical staining was applied to observe the expression of caspase-3 in prostate cancer tissue of mice of the three groups. Results: The qualities and volume increment of tumors in groups B and C were significantly lower than those of group A (P 0.05). Immumohistochemical staining revealed that the caspase-3 protein in prostate cancer tissue of mice of group A expressed negatively with colorless or light-colored karyon; while the caspase-3 protein in prostate cancer tissue of mice of group B expressed positively with dark-colored karyon, centralized distribution and granular sensation; and the caspase-3 in prostate cancer tissue of mice of group C showed strong positive expression with big and darker colored karyon and dense distribution. Conclusions: Oridonin can inhibit the growth of RM-1 prostate cancer cells effectively and have great therapeutic effects on RM-1 cell transplantation tumor of prostate cancer.

Research progress of structural modification of oridonin / 药学实践杂志

Oridonin is an en-kaurene diterpenoids compound which was found to have effect on tumor in vitro.But the clinical application has been limited because of poor water solubility,low bioavailability and oral absorption.Many scholars conducted a large number of structure modification researches of oridonin to search for new diterpenes with higher bio-activities and better bio-availabilities,and to develop new antitumor drugs.In this paper,studies on this aspect of oridonin were re-viewed.

Therapeutic effect of oridonin on mice with prostate cancer

OBJECTIVE@#To investigate the therapeutic effect and the related mechanism of oridonin on mice with prostate cancer.@*METHODS@#Sixty BALB/C male nude mice were selected. A model of RM-1 cell transplantation tumor of prostate cancer was built by the subcutaneous inoculation of RM-1 cells. After that, those 60 experimental mice were randomly divided into groups A, B and C. Each group had 20 mice. Mice in group A were treated with 0.2 mL of normal saline (0.9%) by intraperitoneal injection once a day; mice in group B received intraperitoneal injection of 1.875 mg/mL of oridonin once a day; and mice in group C received intraperitoneal injection of 7.5 mg/mL of oridonin once a day. Mice in the three groups were treated uninterruptedly for 5 weeks and were all killed. Then, tumors were excised and weighed to calculate their growth inhibitory rate, volume increment and anti-tumor rate. Thymus and spleen of mice in the three groups were collected to calculate the thymus and spleen index. Immunohistochemical staining was applied to observe the expression of caspase-3 in prostate cancer tissue of mice of the three groups.@*RESULTS@#The qualities and volume increment of tumors in groups B and C were significantly lower than those of group A (P  0.05). Immumohistochemical staining revealed that the caspase-3 protein in prostate cancer tissue of mice of group A expressed negatively with colorless or light-colored karyon; while the caspase-3 protein in prostate cancer tissue of mice of group B expressed positively with dark-colored karyon, centralized distribution and granular sensation; and the caspase-3 in prostate cancer tissue of mice of group C showed strong positive expression with big and darker colored karyon and dense distribution.@*CONCLUSIONS@#Oridonin can inhibit the growth of RM-1 prostate cancer cells effectively and have great therapeutic effects on RM-1 cell transplantation tumor of prostate cancer.

Effect of dexamethasone combined with oridonin on proliferation and apoptosis of multiple myeloma cell line U266 / 白血病·淋巴瘤

Objective To investigate the effect of dexamethasone combined with oridonin on proliferation and apoptosis in multiple myeloma cells U266 and the related molecular mechanism. Methods Exponential phase of growth U266 cells were treated with different concentrations of oridonin combined with dexamethasone or alone. U266 cells treated by DMSO were taken as control group. The proliferation inhibitory ratios were measured by CCK-8 assay followed by 24 h, 48 h and 72 h. Apoptosis induction was assessed by using Annexin V-FITC kit. Real time PCR was used to examine the mRNA changes of Notch1, NF-κB/p65 and bcl-2. Western blot assay was applied to detect the protein expression of Notch1, cleaved Notch1, NF-κB/p65 and bcl-2. Results Compared with that in control group, proliferation in all the experimental groups was inhibited (P<0.05), and the apoptosis was promoted (P<0.05); especially the combination of dexamethasone and oridonin had a synergistic effect on the proliferation and apoptosis of U266 cells (P<0.05). The results of PCR and Western blot showed that after treatment of U266 cells with dexamethasone, the mRNA as well as their protein levels of NF-κB/p65 and bcl-2 were decreased compared with those in the control group (P<0.05). Moreover, the mRNA and protein expression of Notch1, cleaved Notch1, NF-κB/p65 and bcl-2 was obviously down-regulated in oridonin group and the combination group (P<0.05). Conclusion Combination of dexamethasone and oridonin can significantly increase the anti-tumor effect by inhibiting proliferation and inducing apoptosis of U266 cells, which may be related to the inhibition of the Notch1 pathway.