[Expression of Runt-related Transcription Factor 3 in Human Colon Cancer Cell Line HCT-116 Resistant to 5-Fluorouracil and the Mechanism of Drug Resistance].

Objective To establish a human colon cancer cell line HCT-116/5-FU resistant to 5-fluorouracil(5-FU)and explore the relationship between runt-related transcription factor 3(RUNX3)and drug resistance of colorectal cancer.Methods The human colon cancer cell line HCT-116/5-FU with resistance to 5-FU was established by low concentration gradient increment combined with high-dose intermittent shock.CCK-8 method was used to determine the half maximal inhibitory concentration(IC50)of 5-FU on the parent line HCT-116 and drug-resistant line HCT-116/5-FU.The cell growth curve was established for the calculation of population doubling time(TD).The mRNA levels and protein levels of RUNX3,P-glycoprotein(P-gp),multidrug resistance-associated protein 1(MRP1),and lung resistance-related protein(LRP)in HCT-116 and HCT-116/5-FU cells were determined by qRT-PCR and Western blotting,respectively.The RUNX3 expression in HCT-116 cells was knocked down by siRNA technique,and the cells were divided into RUNX3 knockdown groups(si-RUNX3-1 group and si-RUNX3-2 group)and negative control group(si-NC group).The knockdown efficiency was verified by qRT-PCR at the mRNA level and Western blotting at the protein level.The IC50 in si-RUNX3 groups and si-NC group was determined with CCK-8 method,and the expression of P-gp,MRP1,and LRP in the two groups was detected by Western blotting.Results A stable human colon cancer drug-resistant cell line HCT-116/5-FU was successfully constructed.HCT-116/5-FU showed the TD 1.38 times as long as that of HCT-116(P=0.002)and changed morphology.The mRNA level of RUNX3 in HCT-116/5-FU cells was significantly lower than that in HCT-116 cells(P=0.048),and those of P-gp(P=0.008),MRP1(P=0.001),and LRP(P=0.001)showed the opposite trend.The protein level of RUNX3 in HCT-116/5-FU cells was significantly lower than that in HCT-116(P<0.001),and those of P-gp,MRP1,and LRP presented the opposite trend(all P<0.001).The HCT-116 cell model with low expression of RUNX3 was successfully established.The mRNA level of RUNX3 had no significant difference between si-RUNX3-1 group and si-NC group(P=0.064),while the level in si-RUNX3-2 group was significantly lower than that in si-NC group(P=0.034).The protein levels of RUNX3 in si-RUNX3-1 group and si-RUNX3-2 group were lower than that in si-NC group(both P<0.001).The results demonstrated higher knocking efficiency in si-RUNX3-2 group,which was thus selected to complete the follow-up test.The IC50 of si-RUNX3 group was significantly higher than that of si-NC group(P<0.001),which indicated that the down-regulated expression of RUNX3 could reduce the sensitivity of HCT-116 cells to 5-FU.The relative protein levels of P-gp,MRP1,and LRP in si-RUNX3 group were significantly higher than those in si-NC group(all P<0.001).Conclusion The down-regulation of RUNX3 expression can reduce the sensitivity of HCT-116 cells to 5-FU,which is considered to be related to the up-regulated expression of P-gp,MRP1,and LRP.

[Effect of Acupuncture Therapy on Body Compositions in Patients with Obesity].

OBJECTIVE: To observe the clinical effectiveness of acupuncture intervention in weight reduction by modulating body compositions in obesity patients. METHODS: A total of 71 obesity patients during weight-loss procedure were allocated to acupuncture+nutrition-consultation group (n=40) and simple nutrition-consultation group (n=31). The patients of the acupuncture +nutrition-consultation group were treated by acupuncture stimulation of Zhongwan (CV 12), Xiawan (CV 10), Tianshu (ST 25), Wailing (ST 26), Qihai (CV 6), Guanyuan (CV 4), etc. for 30 min, once every other day, 3 times per week, 12 times altogether, and also given with weekly nutrition consultation (including subjective query, objective measurement, analysis, program for nutrition support) at the same time. The patients of the simple nutrition-consultation group were treated by only weekly nutrition consultation for 4 weeks. Before and after the treatment, the patients' body weight, body mass index (BMI), fat mass, percentage of body fat, muscle mass, protein quality, water quality and bone mass were measured by using a composition analyzer. RESULTS: After 4 weeks' treatment, the body mass, BMI, fat mass and fat percentage in both acupuncture+nutrition-consultation and simple nutrition-consultation groups were significantly decreased (P<0.01), while the weight levels of muscle, protein, bone and water content had no apparent changes (P>0.05). The therapeutic effect of acupuncture+nutrition-consultation group was markedly superior to that of the simple nutrition-consultation group in increasing the improved degrees of body weight, BMI, fat mass and fat percentage (P<0.01). CONCLUSIONS: Acupuncture plus nutrition consultation is effective in reducing body mass, fat mass and percentage of body fat in obesity patients.