| Objective To investigate the effects on the growth, apoptosis, and the express ofFas/FasL in K562, K562/ADM in vitro by nociceptin. Try to discover the mechanism onapoptosis of leukemia cell by nociceptin. Find how nociceptin to reverse the ADMresistance and the mechanism on K562/ADM. Analysis the related effects on MM andCLL by FISH.Methods MTT was used to analysis on the growth of K562 and K562/ADM;Cytometry flow were used to detect the apoptosis of K562 and K562/ADM, theexpression of related genes by nociceptin; Cytometry flow was used to find the changesof P-gp, [ADM], [Ca2+] before and after the treatment by nociceptin; FISH was used toinvestigate the changes on chromosomes of MM and CLL; Analysis the data togetherwith Clinical indicators.Results①The inhibitory effect on the growth of K562 and K562/ADM istime-related. The best effect is caused by 10-9 mol·L-1 of nociceptin and got the highesteffect at 96 hours. 10-7, 10-9 and 10-12 mol·L-1 of nociceptin caused the cytotoxic effectafter 72 hours （P＜0.05）.②After 72 hour treatment of 10-7, 10-7 and 10-9 mol·L-1 ofnociceptin, the highest peak of apoptosis apeared, 22.8%, 23.8%, 26.5% respectively.G0/G1 cell increased, and S cell decreased.③OFQ could reverse the ADM ofK562/ADM. IC50 was 93.3±0.14μg·ml-1, 46.99±0.25μg·ml-1, 35.31±0.26μg·ml-1 respectively after treatment of nociceptin 24, 48, and 72 hours. Afterco-treatment of ADM and non-cytotoxic dose nociceptin (10-7 mol·L-1) 24, 48 and 72hours, IC50 was 70.04±0.19μg·ml-1 （1.33*）, 33.21±0.27μg·ml-1 （1.42*）, 23.11±0.29μg·ml-1 （1.53*） respectively.④The expression of P-gp in K562/S was 1.28%. Itis much lower than the express in K562/ADM 98.23%. After 48 hours treatment bydifferent amount of nociceptin, the express of P-gp in K562/ADM was decreased indose-related mode.⑤After treatment by nociceptin, the accumulation of ADM wasincreased in time-related mode.⑥The FISH results of MM patients showed that,chromosomes changed in 75% (12/16, IgH recomposition is 69% （11/16）; 13q missing（8/16）; RBI is 38% （6/16）; D13S319 is 25% （4/16）; 1q21 increased 38% （6/16）; thechromosomes changed in 3 positions or above is 42%; there is no p53 change.⑦FISH（5 probes） results of 10 CLL patients showed that, there is at least 1 molecular geneticsabnormal in 9 patients; there is no changes found only 1 patient; I of 2 BinetB patientwas found chromosome changes; for 8 BinetC patients, 2 were found abnormal karyotype; 7 were found gene missing; 1 was found gene increasement; 2 were foundhomozygous deletion.Conclusions①Nociceptin could inhibit the growth K562 and K562/ADM in vitro. It istime and dose-belated.②Nociceptin caused apoptosis of K562.③Nociceptin couldreverse the AMD resistance in K562/ADM partially.④Nociceptin could decrease theexpress of P-gp, increase the concentration of ADM in cell, than increase the cytotoxiceffect of chemotherapy drugs.⑤Nociceptin could increase the express of Fas, FasL inK562/ADM. Fas and FasL could cause apoptosis.⑥IGH rearrangement usually happenin the early stage of MM disease with high rates. These are related with high level ofBeta2 microglobulin and are bad factors of prognosis. 1q21 rearragement usuallyhappens in the progression stage of MM disease. 13q deletion happens veryfrequently in chromosomal abnormalities, especially in MM disease. It is also a badfactor in prognosis. 1q21 gene amplification is not primaryily morphological changes. Itusually happens in the 2nd stage of MM disease. It needs further investigations whetherit is related with clinical symptoms of MM disease.⑦Del（13q14） abnormality most oftenhappens in CLL disease. Del（ATM） and del（17q13.1） abnornality shows bad prognosis. If+12 abnornality is accompany with multi-gene locus abnormalities, prognosis is alsobad in CLL disease. |
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