[Effect of heat stress on expression of gp96 in K562 cell line of the chronic myeloid leukemia and its significance].
This study was purposed to investigate the effect of different heat stress conditions on expression level of heat shock protein gp96 in K562 cell line of chronic myeloid leukemia in order to provide experiment basis for seeking optimal heat stress condition increasing extraction amount of gp96 from K562 cells. The expression changes of gp96 in K562 cell line was detected by immunocytochemistry under 38, 40, 42, 44, 46 and 48 degrees C for 30 minutes in water, by flow cytometry under 40, 44, 48 and 52 degrees C for 30 minutes in water, by Western blot under 40, 44, 48 and 52 degrees C for 30 minutes in water. Immunocytochemistry assay showed that gp96 existed mainly in cytoplasm. The peak of gp96 expression was at 30 minutes after 48 degrees C in water. The result of flow cytometry was consistent to immunocytochemistry detection results under temperatures 40, 44, 48 and 52 degrees C (P < 0.01). Western blot showed that detection result was the same as the immunocytochemistry and flow cytometry detections. In conclusion, the expression of gp96 in K562 cell line reached peak at 30 minutes after 48 degrees C in water. This condition may be an effective preparative condition for extraction of gp96 from K562 cells.
Wang
Xin
X
Department of Hematology, Fuxing Hospital, Capital Medical University, Beijing 100038, China.
Shi
Bin
B
Wang
Shen-Wu
SW
Dong
Jian-Qiang
JQ
Cui
Jian-Ying
JY
Han
Shu-Xia
SX
D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D015153:Blotting, Western; D005434:Flow Cytometry; D018869:Heat-Shock Response; D006801:Humans; D007150:Immunohistochemistry; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism* / Q000473:pathology
D000951:Antigens, Neoplasm; C052646:sarcoma glycoprotein gp96 rejection antigens