ABSTRACT
BACKGROUND: The Id2 gene is an endogenous negative regulator of basic helix-loop-helix factor, which is involved in the cell proliferation, differentiation and existence. Id2 also shows functional diversity in the progression and infiltration of different types of tumors OBJECTIVE: To observe the changes of proliferation and invasiveness of PC-3 human prostate cancer stem cells after shRNA-Id2 transfection. METHODS: PC-3 human prostate cancer stem cells in logarithmic growth phase were harvested to isolate tumor stem cell spheres by serum-free suspension culture. The expression of CD44+CD24-on the surface of tumor stem cells was detected by flow cytometry. The shRNA-Id2 expression vector was constructed and transfected into PC-3 human prostate cancer stem cells. Untransfected PC-3 human prostate cancer stem cells were used as control. At 48 hours after transfection, the expression of Id2 gene and protein in shRNA-Id2 transfected prostate cancer stem cells, NC-shRNA empty vector transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by RT-PCR and western blot, respectively. The proliferation and invasion of shRNA-Id2 transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by MTT assay and Transwell chamber, respectively. The expressions of E-cadherin, vimentin and Twist were detected by western blot and RT-PCR. RESULTS AND CONCLUSION: Tumor stem cell spheres were successfully isolated by the serum-free suspension culture. The expression rate of CD44+CD24-on the surface of the third-generation PC-3 human prostate cancer stem cells was (85.69±8.96) ％, indicating that the cultured tumor stem cell spheres overexpressed the phenotype of tumor stem cells. At 48 hours after transfection, the expression of Id2 gene and protein was significantly lower in the shRNA-Id2 transfection group than the non-transfection group (P < 0.05) , indicating that the expression of Id2 was successfully interfered with the expression of PC-3 prostate cancer stem cells. The invasive ability of the cells in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). Western blot and RT-PCR detection showed that the expression of E-cadherin, an epithelial marker of PC-3 prostate cancer stem cells, in the shRNA-Id2 transfection group was significantly higher than that in non-transfection group (P < 0.05) , while the expression of vimentin, a marker of mesenchymal stem cells, and Twist, a transcription factor regulating cell-mesenchymal transformation, in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). These findings indicate that RNA interference with Id2 gene can inhibit the proliferation and invasion of PC-3 prostate cancer stem cells by regulating the expression of E-cadherin, vimentin and Twist.
ABSTRACT
BACKGROUND: The Id2 gene is an endogenous negative regulator of basic helix-loop-helix factor, which is involved in the cell proliferation, differentiation and existence. Id2 also shows functional diversity in the progression and infiltration of different types of tumors OBJECTIVE: To observe the changes of proliferation and invasiveness of PC-3 human prostate cancer stem cells after shRNA-Id2 transfection. METHODS: PC-3 human prostate cancer stem cells in logarithmic growth phase were harvested to isolate tumor stem cell spheres by serum-free suspension culture. The expression of CD44+CD24-on the surface of tumor stem cells was detected by flow cytometry. The shRNA-Id2 expression vector was constructed and transfected into PC-3 human prostate cancer stem cells. Untransfected PC-3 human prostate cancer stem cells were used as control. At 48 hours after transfection, the expression of Id2 gene and protein in shRNA-Id2 transfected prostate cancer stem cells, NC-shRNA empty vector transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by RT-PCR and western blot, respectively. The proliferation and invasion of shRNA-Id2 transfected prostate cancer stem cells and untransfected prostate cancer stem cells were detected by MTT assay and Transwell chamber, respectively. The expressions of E-cadherin, vimentin and Twist were detected by western blot and RT-PCR. RESULTS AND CONCLUSION: Tumor stem cell spheres were successfully isolated by the serum-free suspension culture. The expression rate of CD44+CD24-on the surface of the third-generation PC-3 human prostate cancer stem cells was (85.69±8.96) ％, indicating that the cultured tumor stem cell spheres overexpressed the phenotype of tumor stem cells. At 48 hours after transfection, the expression of Id2 gene and protein was significantly lower in the shRNA-Id2 transfection group than the non-transfection group (P < 0.05), indicating that the expression of Id2 was successfully interfered with the expression of PC-3 prostate cancer stem cells. The invasive ability of the cells in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). Western blot and RT-PCR detection showed that the expression of E-cadherin, an epithelial marker of PC-3 prostate cancer stem cells, in the shRNA-Id2 transfection group was significantly higher than that in non-transfection group (P < 0.05), while the expression of vimentin, a marker of mesenchymal stem cells, and Twist, a transcription factor regulating cell-mesenchymal transformation, in the shRNA-Id2 transfection group was significantly lower than that in the non-transfection group (P < 0.05). These findings indicate that RNA interference with Id2 gene can inhibit the proliferation and invasion of PC-3 prostate cancer stem cells by regulating the expression of E-cadherin, vimentin and Twist.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Shao Yao-Gan Cao-Tang on function and expression of P-glycoprotein (P-gp) in Caco-2 cells.</p><p><b>METHOD</b>3H-digoxin (Dig), a substrate of P-glycoprotein, was used as a probe to measure the P-gp-mediated drug efflux transport, which indicated the function of P-gp in Caco-2 cells, while Verapamil (Ver) was used as a positive P-gp inhibitor. P-gp expression in Caco-2 cells was tested by immunohistochemistry staining. Inhibition effect of SGT on P-gp-mediated drug efflux transport and P-gp expression were investigated.</p><p><b>RESULT</b>Dig was shown a positive absorption mode in Caco-2 cell monolayer, characterized as the ratio of apparent permeabilities (Papp) from basolateral side to apical side Papp (BL-->AP) and from apical side to basolateral side Papp (AP-->BL) of Dig was 27.07. Addition of Ver into Dig transport media significantly inhibited P-gp activity which was indicated by increasing the Papp (AP-->BL) of Dig by 3.82 times, whereas Ver had no significant effect on Papp (BL-->AP). SGT (at the concentrations of 1/25 IC5, 1/5 IC5, IC,) could promote Papp (AP-->BL) of Dig by 159.83%, 217.95% ,160.26%. Papp (AP-->BL) of Dig was mildly increased by 59.16%, 50.73% by SGT at 1/25 IC5, 1/5 IC, respectively. Immunohistochemistry staining showed that SGT inhibited the expression of P-gp of Caco-2 cells.</p><p><b>CONCLUSION</b>SGT showed the potential inhibition to the function and expression of P-gp, leading to the increase absorption of P-gp's substrates.</p>
Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B , Metabolism , Biological Transport , Caco-2 Cells , Drug Interactions , Drugs, Chinese Herbal , PharmacologyABSTRACT
Potential drug interactions on P-glycoprotein transport level were discussed in this article. As an efflux transporter protein widely distributed in various tissues, P-glycoprotein plays an important role in many drugs interactions, via affecting the metabolic process to alter the concentration of drug in plasma and tissue. Interactions between herb and western drug may occur by inducing or inhibiting P-glycoprotein, resulting in either efficacy enhancement or adverse effects. So it is important for clinical drug use by rational utilization of the interaction between herb and western drug which medicated by P-glycoprotein.
Subject(s)
Animals , Humans , ATP Binding Cassette Transporter, Subfamily B , Metabolism , Drug Interactions , Drugs, Chinese Herbal , Pharmacology , Protein TransportABSTRACT
In the treatment of 63 cases of depression, by bilateral Baihui (GV 20),Sishencong (Ex-HN 1), Juque (CV 14) and Neiguan (PC 6) as the main acupoints, based upon pattern identification, with bilateral Zhigou (TE 6), Zusanli (ST 36), Yanglingquan (GB 34)and Taichong (LR 3) added for liver qi stagnation and spleen deficiency, with bilateral Hegu (LI 4), Xuehai (SP 10), Sanyinjiao (SP 6) and Taichong (LR 3) added for stagnation of liver blood, with bilateral Shenmen (HT 7), Zusanli (ST 36), Sanyinjiao (SP 6) and Taibai (SP 3)added for deficiency in both the heart and spleen, and with bilateral Sanyinjiao (SP 6), Taibai(SP 3), Taixi (KI 3) and Guanyuan (CV 4), by moxibustion on Guanyuan (CV4) and needling techniques on the rest acupoints, the results showed clinical cure in 21 cases, remarkable effect in 18 cases, improvement in 20 cases and failure in 4 cases, in the treatments from 13times to 45 times, at the average treatments of 26 times.
ABSTRACT
BACKGROUND:Attention deficit hyperactivity disorder(ADHD) is treated clinically mainly with medication,psychotherapy and acupuncture,which is seldom treated with simple massage.During working in Swaziland, the writer found a part of children only accept massage for they did not receive remarkable results by medication and also were scared of acupuncture. OBJECTIVE:To probe manipulation and mechanism of massage on ADHD in children. DESIGN:Cases analysis. SETTING:Sino Japan Friendship Hospital. PARTICIPANTS:All the cases were collected from April 2000 to April 2002 while the writer was working in Swaziland. Thirty three children with ADHD were in accordance with the symptomatic diagnostic standards of American DSM IV.No control was selected. INTERVENTIONS:Simple massage manipulation was adopted in treatment of the 33 ADHD children.The common manipulations included head manipulation,chest abdominal manipulation and back manipulation.According to various clinical manifestations,the manipulations were modified based on differentiation of syndromes. MAIN OUTCOME MEASURES:Therapeutic results on patients with massage manipulations. RESULTS:Of the 33 cases of ADHD children treated with massage, 10 were cured,9 were remarkably effective,9 were improved and 5 had no effect. CONCLUSION:The therapeutic results of massage on ADHD in children are satisfactory and safe without side effects.
ABSTRACT
The main acupoints ofZhongwan (CV 12), Tianshu (ST 25), Zusanli (ST 36), Shangjuxu (ST 37), Dachangshu (BL 25) and Shenque (CV 8), in combination with the corresponding acupoints based on the syndrome differentiation, were selected to treat 28 cases of chronic nonspecific ulcerative colitis, the result showed clinical cure in 18 cases, clinical effect in 15 cases, failure in 5 cases. The most treatment times were 46, and the least 25.