ABSTRACT
Objective:To investigate the effect of actin related protein 2/3 complex subunit 2 (ARPC2) gene silencing on the biological characteristics of papillary thyroid carcinoma (PTC) TPC-1 cells through lentivirus-mediated RNA interference.Methods:TPC-1 cells infected with nonsense short hairpin RNA (shRNA) sequence lentivirus (shCtrl) was used as the control group. TPC-1 cells infected with ARPC2 shRNA interference sequence lentivirus (shARPC2) was used as the experimental group, in which the expression of ARPC2 gene was specifically interfered. The effects of silencing the expression of ARPC2 gene on the proliferation of TPC-1 cells were detected by using methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, Western blot and colony formation test. Flow cytometry and Western blot were conducted to detect the effect of silencing ARPC2 gene on TPC-1 cells apoptosis and related proteins.Results:shARPC2 could efficiently infect TPC-1 cells, and the expression efficiency of green fluorescent protein was over 85%. Compared with the control group, TPC-1 proliferation was inhibited in the experimental group. The ratio of S-phase cells in the experimental group was reduced compared with that in the control group [(14.79±0.21)% vs. (21.13±0.33)%, t = 27.77, P < 0.05]. The ratio of G 1 and G 2/M-phase cells in the experimental group was increased compared with that in the control group [G 1 phase: (67.57±0.08)% vs. (62.06±0.36)%, t=25.56, P < 0.05; G 2/M phase: (17.64±0.12)% vs. (16.91±0.17)%, t=6.154, P < 0.05]. Meanwhile, the expressions of cell cycle-related proteins CDK2, CyclinE and CyclinD were reduced in the experimental group. The number of clone formation in the experimental group was less than that in the control group, the difference was statistically significant [(10±2) vs. (161±6), t=9.011, P < 0.05]. In addition, the apoptotic ratio of cells in the experimental group was higher than that in the control group [(8.60±0.77)% vs. (4.08±0.40)%, t=9.011, P < 0.05]. Western blot showed that the expressions of anti-apoptotic factors p21 and bcl-2 were reduced in the experimental group, while the expression of pro-apoptotic factor bax was increased. Conclusion:The interference with the expression of ARPC2 regulated by shRNA can inhibit the proliferation, and promote the apoptosis of PTC TPC-1 cells, indicating that ARPC2 may be a possible biological new target for the treatment of PTC.
ABSTRACT
Objective To investigate the surgery-related complications and risk factors of ileocolic Crohn's disease (CD).Methods The retrospective case-control study was conducted.The clinicopathological data of 52 patients with ileocolic CD who underwent surgery at the Peking Union Medical College Hospital from January 2010 to April 2016 were collected.Observation indicators:(1) surgery-related complications,(2) risk factors analysis of surgery-related complications:gender,age of onset,preoperative body mass index (BMI),course of disease,smoking history,history of appendectomy,perianal lesions,oral ulcer,C-reactive protein (CRP),erythrocyte sedimentation rate,disease behavior,short crohn's disease activity index (sCDAI),preoperative amino salicylic acid therapy,preoperative hormone therapy,preoperative antituberculosis therapy,preoperative immunosuppressive agents therapy,preoperative biologic agents therapy,emergency operation,surgical method and ileocolic anastomosis method,(3) follow-up.The follow-up using outpatient examination and telephone interview was performed to detect recurrence of disease up to August 2016.Measurement data with normal distribution were represented as-x ± s.The univariate analysis was done using the chi-square test,and multivariate analysis was done using the Logistic regression model.Results (1) Surgery-related complications:of 52 patients,12 had postoperative complications.Four patients complicated with wound infection had good healing of the wound after debridement and dressing change.Of 4 patients with abdominal infection,3 were improved by anti-infection symptomatic treatment and 1 die of septic shock at postoperative day 1.One patient with intestinal obstruction had a smooth recovery after open adhesiolysis.One patient with intestinal fistula discharged from hospital due to a critical condition under families' requestion.One patient with acute cholecystitis and 1 with acute pancreatitis were respectively improved by conservative treatment.(2) Risk factors analysis of surgery-related complications:theresult of univariate analysis showed that sCDAI and emergency operation were the factors infecting surgery-relatedcomplications of ileocolic CD (x2 =6.299,8.494,P ＜ 0.05).The result of multivariate analysis showed that sCDAI was an independent factor infecting surgery-related complications of ileocolic CD [OR =2.716,95％ confidence interval (CI):1.216-6.066,P ＜ 0.05].(3) Follow-up:all the 52 patients were followed up for 5-76 months with a median time of 39 months.During the follow-up,15 had recurrence of diseases and then underwent medical treatment.Conclusions Patients with ileocolic CD are easily complicated with wound infection and abdominal infection in the active period,and sCDAI is an independent factor infecting surgery-related complications of ileocolic CD.