The Inhibitory Effect Of Hyperthermia Combined With Cisplatin On The Proliferation Of Human Gastric Cancer Drug- Cells SGC-7901/DDP And Its Molecular Mechanism

Objective:To research the effects of hyperthermia?HT?combined with cisplatin?DDP?on the inhibition of human gastric cancer drug-resistant cell SGC-7901/DDP in vitro and explore its possible molecular mechanism.To provide an evidence for the research and treatment of cisplatin combined with hyperthermia in patients with gastric cancer resistance.Methods:Routine culture tumor resistant cell SGC-7901/DDP in vitro.Divide cells into control group,cisplatin group,hyperthermia group and hyperthermia combined with cisplatin group.Observe cell morphology with inverted phase contrast microscope after being cultured at 41?,44?,47? and 50? for 12 h,24 h,36 h;MTT assay detected the proliferation of SGC-7901/DDP at different time and temperature,and the proliferation of SGC-7901/DDP at 41?,44? and 47? combined with 1 ?g/ml,2 ?g/ml and 3 ?g/ml cisplatin concentrations,to clarify the interaction between hyperthermia and cisplatin;Using Annexin V-FITC/PI double staining assay apoptotic rate of SGC-7901/DDP cells in control group,cisplatin group?2 ?g/ml?,hyperthermia group?47??,and cisplatin combined hyperthermia group?47? and 2 ?g/ml?.High-throughput chip technique was applied to test lncRNAs expression in SGC-7901/DDP cells from the control group,cisplatin group and hyperthermia combined with cisplatin groups.Then,to verify the expression of lncRNA TCONS₀0018082,TCONS₀0015171,ENST00000584911.1,ENST00000412526.1,and ENST00000592460.1 in cisplatin combined hyperthermia group versus control group by real-time fluorescence quantitative polymerase chain reaction?RT-PCR?.Results:?1?MTT results showed that the inhibition of SGC-7901/DDP cell proliferation was dramatically enhanced after 47? hyperthermia combined with 2 ?g/ml cisplatin compared with the single factor intervention?P<0.05?and the control group?P<0.05?,and the combination had a synergistic effect;?2?Under the same conditions,Annexin V-FITC/PI double staining experiment result expressed that hyperthermia combined with cisplatin increased the rate of early apoptosis of SGC-7901/DDP cells compared with using the hyperthermia or cisplatin alone;?3?LncRNA high-throughput chip expression profile showed that a large number of lncRNAs and mRNAs opposite to the control group appeared in the cells of the hyperthermia combined cisplatin group;?4?RT-PCR test results confirmed that lncRNA TCONS₀0018082 and ENST00000412526.1 were significantly up-regulated?P<0.01?,while the relative expression levels of lncRNA TCONS₀0015171 and ENST00000584911.1 were significantly down-regulated?P<0.01?.Conclusions:?1?After being cultured for 24 hours,the hyperthermia at 47? combined with 2 ?g/ml cisplatin can synergistically inhibit human gastric cancer drug-resistant cells SGC-7901/DDP proliferation and promote early apoptosis;?2?Hyperthermia at 47? combined with 2 ?g/ml cisplatin reversed the expression of a large number of mRNA and lncRNA in human gastric cancer drug-resistant cells SGC-7901/DDP.?3?The molecular mechanism of inhibiting proliferation of human gastric cancer drug-resistant cells SGC-7901/DDP by combining hyperthermia at 47? with 2 ?g/ml cisplatin may be closely related to up-regulation of TCONS₀0018082 and ENST00000412526.1,and down-regulation of TCONS₀0015171 and ENST00000584911.1.