ABSTRACT
The purpose of this research is to investigate the effects and mechanisms of paeonol (PL), a phenolic compound found in many traditional Chinese formulations, on reversing drug resistance in the ovarian cancer resistant SKOV3/DDP cells. The results showed that PL had significant drug-resistant reversal effect on SKOV3/DDP cells. Flow cytometry showed that PL could inhibit P-glycoprotein (P-gp) function in a concentration-dependent manner. Fluorescent quantitative PCR and cell immunofluorescence techniques were used to detect mechanisms of action. Results revealed that both the inhibitory effect on MDR1/P-gp and metadherin (MTDH) expression and the induction effect on phosphatase and tensin homolog (PTEN), by 15, 30, and 60 μmol·L-1 PL, were increased with increased concentrations of PL (P P -1 PL treated group; however, the inhibition or induction effect on MTDH or PTEN protein were only comparable to the 15 μmol·L-1 PL treated group. The present study shows that the effect of PL on SKOV3/DDP cells may be related to the inhibition of P-gp function and expression, the inhibition of MDR1, MTDH expression, and the induction of PTEN expression, all which can provide a theoretical foundation for PL as a drug resistance reversal agent on the treatment of ovarian cancer chemotherapy resistance.
ABSTRACT
OBJECTIVE To detect the reversal effect of Guizhi Fuling Wan on cisplatin-resistant ovarian cancer SKOV3/DDP cells and its relationship with protein expression of phosphatase and tensin homolog (PTEN) and metadherin (MTDH). METHODS Guizhi Fuling Wan (GFW) concentrated solution was prepared according to the Chinese Pharmacopoeia 2015 edition, Wistar rats were given GFW viagavage at 4 g·kg-1·d-1,8 g·kg-1·d-1,16 g·kg-1·d-1,or given saline as blank control for 5 days.Blood samples were taken and the corresponding drug-containing low-dose sera, medium-dose sear, high-dose sera and blank sera were prepared.The XCELLigence RTCA S16 real-time label-free cell analyzer was used to detect the reversal effect by the sera combined with cisplatin or paclitaxel in SKOV3/DDP cells. Annexin V-FITC and PI double-staining were used to detect the apoptosis-inducing effect of the sera in the cells. RT-qPCR and western blot were used to detect the mRNA and protein expression of PTEN and MTDH after the cells treated with the sera. RESULTS The inhibition rate of low-dose sera against SKOV3/DDP cells was less than 5%.After the low-dose sera combined with cisplatin or pacli-taxel, the IC50 of SKOV3/DDP cells against cisplatin and paclitaxel decreased by 3.01 and 1.79-fold, respectively.The total apoptosis rates induced by the low-dose sera,medium-dose sear,high-dose sera and blank sera in SKOV3/DDP cells were 11.08±0.13,19.42±0.30,24.23±0.31,and 3.21±0.24,respec-tively; there was a significant difference between the groups (P<0.01). RT-qPCR results showed that, compared with the blank serum, the sera can up-regulate the expression of PTEN mRNA and down-regulate the expression of MTDH mRNA in a dose-dependent manner. Western blot results showed that the induction effect to PTEN protein and the inhibition effect to MTDH protein by the sera were gradually enhanced with thesera dose increasement. CONCLUSION The resistance reversal effect of Guizhi Fuling Wan on ovarian cancer SKOV3/DDP cells may be related to the inhibition of MTDH, up-regulation of PTEN and induction of apoptosis, providing with an experiment basis for the applica-tion of Guizhi Fuling Wan as a reversal agent for chemotherapy resistance of ovarian cancer.
ABSTRACT
Objective To obtain the three-dimensional (3D) motion space of ankle by an integral for the 3D rotation angle with mutual relationships of rotation angles in different directions, which is used to evaluate the flexibility and recovery of ankle more accurately and ful-ly. Methods Twenty-five graduate students were recruited to participate in this study, one group consisted of 20 healthy individuals without ankle injury, and the other was composed of 5 patients with ankle injury. A motion capturing system was used to simultaneously capture the 3D coordinates of the marked points on the foot. Next, these 3D coordinates were converted into rotation angles through trigonometric func-tions processed. The maximum rotation angles of adduction, abduction, varus, and eversion change with the rotation angles of plantar flex-ion and dorsal flexure were measured by using polynomial fitting. At last, the size of the ankle's 3D motion space was obtained by integrat-ing the fitting functions. Results The ankle's 3D motion space of the healthy people was 41.256, and the variation among them was less than 10%. The ankle's 3D motion space of the patient side was 33.163, and the variation among them was less than 15%because of the different degrees of rehabilitation. There was significant difference between two groups (t=8.804, P<0.01). With the same rotation angles of plantar flexion or dorsal flexure, the maximum rotation angle of varus was larger than that of eversion, and the maximum rotation angle of adduc-tion was larger than that of abduction. Conclusion The algorithm can be used to quantify the 3D motion space of ankle more accurately, and to achieve a more comprehensive evaluation of ankle's flexibility and rehabilitation.
ABSTRACT
Objective To obtain the three-dimensional (3D) motion space of ankle by an integral for the 3D rotation angle with mutual relationships of rotation angles in different directions, which is used to evaluate the flexibility and recovery of ankle more accurately and ful-ly. Methods Twenty-five graduate students were recruited to participate in this study, one group consisted of 20 healthy individuals without ankle injury, and the other was composed of 5 patients with ankle injury. A motion capturing system was used to simultaneously capture the 3D coordinates of the marked points on the foot. Next, these 3D coordinates were converted into rotation angles through trigonometric func-tions processed. The maximum rotation angles of adduction, abduction, varus, and eversion change with the rotation angles of plantar flex-ion and dorsal flexure were measured by using polynomial fitting. At last, the size of the ankle's 3D motion space was obtained by integrat-ing the fitting functions. Results The ankle's 3D motion space of the healthy people was 41.256, and the variation among them was less than 10%. The ankle's 3D motion space of the patient side was 33.163, and the variation among them was less than 15%because of the different degrees of rehabilitation. There was significant difference between two groups (t=8.804, P<0.01). With the same rotation angles of plantar flexion or dorsal flexure, the maximum rotation angle of varus was larger than that of eversion, and the maximum rotation angle of adduc-tion was larger than that of abduction. Conclusion The algorithm can be used to quantify the 3D motion space of ankle more accurately, and to achieve a more comprehensive evaluation of ankle's flexibility and rehabilitation.