The Clinical Study On Solid Tumor By Low-Frequency Ultrasound And The Reversal Of Multidrug Resistance By Low-Frequency Ultrasound And Minyaojian In Vitro

Objective: To investigate the biological dynamics and safety of the CO2microbubble from 5% sodium bicarbonate(NaHCO3),vitamin C and 706 in vivoand vitro, the reversal effect of low-frequency ultrasound on the multi-drugresistance in K562/ADM and MCF-7/ADM and its possible mechanisms and alsowant to evaluate the effect of low-level ultrasound(US) on advanced solid tumor.Methods: The CO2 from 5% sodium bicarbonate,vitamin C and 706 wasobserved by microscope. The CO2 produced in intra-hepatic vessel and portalvessel was studied by Doppler ultrasound. We divided 48 patients into threegroups:A,B and C groups. Twenty patients in A group have ≥2 neoplasms;onegroup(A1) was treated by ultrasound and chemotherapy, the control group(A2)was treated by chemotherapy. Seventeen patients in B group were treated byultrasound treatment, and eleven patients in C group were treated bychemotherapy as control group of B. We evaluated the efficacy and side-effectafter 2 therapy circles and did contrast ultrasonography in 12 patients. Thechange of tumor microvascularity before and after cavitation treatment wasobserved by ultrasound angiography. The effect of non-cytotoxic dose ofultrasound and minyaojian and the IC50 of ADM acted on K562/ADM andMCF-7/ADM was assessed by mehyl-hazol-etrazolinum (MTT)assay. Theexpression of p-gp was assessed by immunohistochemistry(IHC). MDR1 mRNAwas assessed by reversal transcription polymerase chain reaction(RT-PCR). Theapoptosis and concentration of intracellular drug were observed by flowcytometry(FCM). Results: After adding 706, the quantity of CO2 significantlyincreased, the diameter of CO2 is similar to that of red cell and the stability ofCO2 improved. The CO2 microbuble is safe and could arrive in intra-hepaticvessel and portal vessel through lung cycle. The signs of enchantment of contrastultrasonography responded less after cavitation treatment. As for efficacy,A1group is better than A2 group, B group is better than C group;but there was notsignificantly difference. The side-effect was higher in C group than B group.Comparing B with C, the quality of life in B group improved significantly. Aftercavitation treatment, 13 cases had PR, and the total effective rate was35.14%(13/37). And the signs level enchantment of contrast ultrasonographyresponded less to the cavitation treatment. MTT assay showed that the inhibitoryrate of K562/ADM and MCF7/ADM in US and US+CO2 group wassignificantly higher than the control group. Reverse multiple of US and US+CO2on K562/ADM was 2.34 and 3.97 respectively. Reverse multiple of US andUS+CO2 on MCF-7/ADM was 2.17 and 3.03 respectively. FCM suggested thatthe administration of low-frequency ultrasound combined with ADMsignificantly increased the apoptosis of K562/ADM and MCF7/ADM. Afterultrasound treatment, the expression of MDR1mRNA and P-gp had nosignificantly change. The noncytotoxic dose of minyaojian was 10ug/ml.MTTassay showed that the inhibitory rate of K562/ADM and MCF7/ADM cell inminyaojian (F6,G2,G3) group was significantly higher than control group.Reverse multiple of F6,G2 and G3 on K562/ADM was 4.31,2.48 and 3.99respectively. Reverse multiple of F6,G2 and G3 on MCF-7/ADM was 2.06,1.58and 2.43 respectively.Rh123 retention test suggested that the administration ofmiyaojian combined with ADM increased the intracellular ADM concentration.After 10ug/ml minyaojian treatment,the expression of MDR1mRNA had nosignificantly change. Conclusions: The CO2 microbubble from 5% sodiumbicarbonate,vitaminC and 706 is a high quality contrast agent: its diameter issimilar to that of red cell, it is safe and could go through lung cycle, and enhanceanti-tumor effect of low-frequency ualtrasound, which is worthy of wide spreadin clinical application.Low-frequency ultrasound is able to enhance thecytotoxicity of ADM and partially reverse the ADM resistance of K562/ADMand MCF7/ADM. The mechanism maybe increase the permeability of cellmember and the concentration of intracellular ADM, promote the apoptosis ofK562/ADM and MCF7/ADM;but there was no effect on the mdr1 gene and p-gpexpression level. Minyaojian is able to enhance the cytotoxicity of ADM andpartially reverse the ADM resistance of K562/ADM and MCF-7/ADM, whichmay be relate to the inhibition of P-gp activity.The efficacy of cavitationtreatment is no less than system chemotherapy, combined with chemotherapy canraise the efficacy. Cavitation treatment can improves the quality of life of patientsand has low even no side-effects. The mechanism of cavitation treatment maybedestroy the tumor vessel. Ultrasound angiography can clearly indicate the changeof tumor microvascularity before and after cavitation treatment and could be usedin clinical tumor therapy.