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目的研究康莱特联合顺铂对骨肉瘤细胞的杀伤作用及其相关机制,探寻骨肉瘤的中西医结合化疗方案。方法免疫组化及RT-PCR方法检测不同浓度康莱特作用下骨肉瘤OS732细胞的Fas表达,并将康莱特与顺铂单独及联合应用于OS732细胞,采用MTT法检测细胞毒性作用,流式细胞仪定量分析凋亡细胞所占比例,相差显微镜及荧光显微镜下观察凋亡细胞形态改变。结果康莱特可上调OS732细胞的Fas表达并诱导凋亡,10μL/mL康莱特与1μlg/mL顺铂合用于OS732细胞的抑制率与各自单用时相比明显提升(P<0.01)。细胞形态观察及凋亡率测定也显示,康莱特与顺铂联用可诱导更多OS732细胞凋亡。结论康莱特可协同顺铂高效杀伤骨肉瘤细胞,这可能与康莱特上调Fas表达诱导骨肉瘤细胞凋亡有关。
Objective To study the killing effect of Kanglaite combined with cisplatin on osteosarcoma cells and its related mechanism and explore the combination of traditional Chinese and Western medicine in the treatment of osteosarcoma. Methods The expression of Fas in osteosarcoma OS732 cells was detected by immunohistochemistry and RT-PCR. KLT and cisplatin alone and in combination were applied to OS732 cells. The cytotoxicity was detected by MTT assay. Flow cytometry The proportion of apoptotic cells was quantitatively analyzed. The morphological changes of apoptotic cells were observed under a phase contrast microscope and a fluorescence microscope. Results KLT could up-regulate the expression of Fas in OS732 cells and induce apoptosis. The inhibitory rates of KLT combined with 10 μL / mL KLT and 1 μM cisplatin on OS732 cells were significantly increased (P <0.01). Cell morphology observation and apoptosis rate also showed that Kanglaite combined with cisplatin can induce more OS732 cell apoptosis. CONCLUSION: KLT can effectively kill osteosarcoma cells in combination with cisplatin, which may be related to KLT up-regulating the expression of Fas and inducing osteosarcoma cell apoptosis.