The Study Of Ultrasound Destructing Taxol Carrying Liposome Microbbles Treatment Of Cervical Cancer HeLa Cell Line

PARTⅠTHE CONCENTRATION OF PACLITAXEL IN CERVICAL CANCER HELA CELLS IN INHIBITORY EFFECTS AND MICROBUBBLES SECT ULTRASONIC RADIATION DAMAGE PARAMETERSObjective: To study the paclitaxel concentration of cervical cancer cells HeLa in inhibitory effects and ultrasonic radiation parameters of destructing microbubbles.Methods: Cell strain was cultured in vitro. The inhibitory effects of different dosages of paclitaxel on cells proliferation and the acute toxic effects of different irradiation time of ultrasound combined blank microbubbles group on cells was detected by MTT assay.Results:1.The proliferation inhibition ratio of HeLa cells in the 72nd hour were determined as (32.37±1.12)%,(42.97±2.18)%,(51.45±0.86)%,(62.48±0.95)%,(69.81±0.57)%,and(76.06±2.4)% after paclitaxel treatment with different concentrations of 10^-9,10^-8,10^-7,10^-6,10^-5 and 10-4mol/L, respectively. And the inhibition ratio of HeLa cells in the 24th hour, 48th hour and 72nd hour were respectively (11.52±1.08)%,(24.6±0.35)%,(32.37±1.12)%when the palictaxel concentration was fixed in 10-9mol/L. The inhibition effect was positively related with the time and concentration of drug treatment, which showed an obvious dose-response time relationship.2. PLM could be destructed with an acoustic intensity of 0.5W/cm2 and the irradiation time of 20 S. When the acoustic intensity was fixed in 0.5W/cm2, and the time of ultrasound radiation on PLM were 10 S, 20 S and 30 S, the survival rates of cells were determined respectively as (92.84±0.66)%,(85.69±2.67)% and (76.61±2.74)%.Conclusion: paclitaxel could inhibit HeLa cells in vitro. There is a little acute toxic effects on HeLa cells using ultrasound combined blank microbubbles with suitable ultrasonic irradiation parameters. Suitable ultrasonic irradiation parameters is 0.5W/cm2, 20 S, and paclitaxel dosage is 10-7mol/L. PARTⅡTHE EFFECT OF PROLIFERATION INHIBITION AND APOPTOSIS INDUCTION ON CERVICAL CANCER HELA USING PACLITAXEL-CARRYING ULTRASOUND MICROBUBBLESobjective: To study the effects of ultrasound combined paclitaxel-carrying liposome microbubbles(PLM) method on HeLa cell proliferation, apoptosis and changes of cell cycle. Methods: 1.HeLa cell strain was cultured in vitro, and divided into 5 groups: paclitaxel group,paclitaxel plus ultrasound irradiation group,paclitaxel- carrying microbubbles plus ultrasound irradiation group ,paclitaxel-carrying microbubbles group and control group. Proliferation inhibition of cervical cancer HeLa cell lines in different groups were observed at different time points by MTT assay. 2. Apoptosis induction of HeLa cell lines in different groups were observed by transmission electron microscope (TEM). 3. The apoptosis of cells and the changes of cell cycle were determined by flow cytometry(FCM).Results:1.Ultrasound combined PLM, paclitaxel alone and ultrasound combined PLM all had significant inhibition effects on HeLa cell proliferation. However, among the three groups, the effects of ultrasound combined PLM group was the strongest (P<0.01). And PLM alone with the concentration equal with the first group had no significant effects on cells.2. Apoptotic bodies were observed by TEM in the ultrasound combined PLM group, apoptotic cells were observed in the ultrasound combined paclitaxel group, and the early apoptotic cells were observed in the paclitaxel alone group. Cell shape in the contrast group and the PLM alone group were both normal.3.The apoptosis rates of cells determined by FCM were (2.18±0.11)% in the contrast group, ( 5.21±0.34 ) % in the PLM alone group, (11.77±0.54)% in paclitaxel alone group, (15.69±0.43)% in the ultrasound combined paclitaxel group and(27.5±0.59)% in the ultrasound combined PLM group, respectively. With the treatment of ultrasound combined PLM, paclitaxel alone and ultrasound combined PLM, the HeLa cells were all arrested in G2/M phase , and with a concurrent decrease in the proportion of S and G0/G1 phase cells .The arreste rates of cells in G2/M phase determined by FCM were (9.7±1.81)% in the contrast group, (14.05±0.8)% in the PLM alone group, (22.04±1.51)% in paclitaxel alone group, (33.28±2.92)% in the ultrasound combined paclitaxel group and (68.28±2.62)% in the ultrasound combined PLM group, respectively.Conclusion: Cervical cancer HeLa cells with paclitaxel-carrying microbubbles have remarkable proliferation inhibition after ultrasound irradiation. HeLa cells is induced aptosis and all arrested in G2/M phase as well .