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Effects of Bone Marrow Fibroblastoid Stromal Cell on the Proliferation and Sensitivity to Chemotherapy of Multiple Myeloma Cell RPMI8226

Author: JinXueQin
Tutor: BaiQingXian
School: Fourth Military Medical University
Course: Internal Medicine
Keywords: Bone marrow microenvironment Multiple Myeloma Bone marrow stromal fibroblast -like cells RPMI8226 cells Joint training across the room Proliferation Resistant cells
CLC: R733.3
Type: Master's thesis
Year: 2004
Downloads: 39
Quote: 0
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Abstract


[Background] Multiple myeloma is a malignant B cells occurred in the hematopoietic system tumors, traditional chemotherapy may initially have some reaction, but eventually the drug-resistant tumor cells. Myeloma cells settle in the bone marrow, peripheral blood and almost not present in other tissues and organs, suggesting that in which the bone marrow microenvironment and its closely related. Bone marrow stromal cells of the bone marrow microenvironment as one of the main ingredients for the increasing influence of myeloma cells caused researchers. Studies have shown that bone marrow stromal cells by means of a variety of cell surface adhesion molecules through ligand - receptor manner combined with myeloma cells, is conducive to tumor cells settle in the bone marrow and produce interactions. However, bone marrow stromal cells on myeloma cells affected by direct or indirect means, or both combined effect has not been fully determined. At home and abroad more research focuses on both bone marrow stromal cells after adhesion by producing IL-6 in myeloma cell proliferation and impact resistance, as other modes of action on how the impact of myeloma cells, needs further discussed. In this study, we use non-IL-6-dependent human myeloma cell line RPMI8226 as target cells, the normal human bone marrow stromal fibroblast-like cell line RPMI8226 cells were cultured with HFCL, HFCL after observing their interaction cell adhesion RPMI8226 cell proliferation and co-cultured with RPMI8226 cells to chemotherapeutic drug sensitivity changes. And the use of newly developed cross-chamber co-culture techniques (Transwell coculture), the two types of cells to further explore ways to do. Fourth Military Medical University, master's degree thesis [Objective 1 observe normal human bone marrow stromal fibroblast-like cells HFCL RPMI8226 myeloma cell proliferation and chemosensitivity. [Methods] RPMI8226 cells and establish direct contact with HFCL cells and indirect contact (Transwen group) co-culture system to RPMI8226 cells cultured alone as the control group. MTT assay HFCL cells and RPMI8226 cell adhesion rate; trypan blue staining was measured RPMI8226 cell growth curve; flow cytometry (FCM) to detect cell cycle changes; Wright's Giemsa staining after a mitotic index (Ml ) determination; Western Blot Detection RPMI8226 cells proliferating cell nuclear antigen (PCNA) expression; MTT assay RPMI8226 cells with different sensitivity to cytotoxic drugs. [Results] ① RPMI8226 cells in direct contact with HFCL cells were incubated for 1 hour, the adhesion between the rate of 29.4%; ② Compared with the control group, the growth curve shows direct contact group RPMI8226 cell growth inhibition, rather than direct contact with the group (Transwell group) did not change: ③ After 72 hours of incubation, the cell cycle by flow cytometry results showed: the control group Gl phase cells accounted for 33.6%, S phase accounted for 52.1%: Direct contact group Gl phase cells increased to 40.4%, S phase cells decreased to 40.8%, compared with the control group is significantly different (P lt; 0.01); Transwell group GI phase cells accounted for 30.6%, S phase cells accounted for 49, l%, compared with the control group, t dagger was no significant difference (P gt; 0.05); ④ culture l, 2,3,4,5 days, mitotic index showed that the control group were: 1.4%, 3.4%, 3%, 1.8%, 1.6%; were cultured in direct contact with HFCL group 1.2%, 2.6%, 2%, 0.8%, 0.4%, lower than the control group: ⑤ cultured for 72 hours, Westem Blot test results show: HFCL direct contact with RPMI8226 cells compared with the control group of proliferating cell nuclear antigen ( PCNA) downregulated. ⑥ with HFCL were cultured for 72 hours after the dose-response curves show: RPMI8226 cells to vincristine, mitoxantrone onions awake, doxorubicin, topotecan reduced sensitivity. [Conclusion 1 ① normal human bone marrow stromal cell line HFCL fibroblasts through direct contact with RPMI8226 can inhibit the proliferation of myeloma cells, preventing the operation of the cell cycle: but with Transwen inserts to divide them after no significant effect on proliferation. ② HFCL fibroblast-like stromal cells and myeloma cells were cultured in direct contact RPMI8226, RPMI8226 cells can make a variety of enhanced tolerance to chemotherapy drugs.

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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Bone marrow tumor
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