Effects of Tetrastigma hemsleyanum Total Extraction on Human γδT Cells Function
XU Qing1, LUO Xiaomei1, YANG Yang2, SUN Xiangxiang1, FENG Liya3, GUO Miaomiao3, LIU Junquan1,3*
The purpose of this paper was to study the effect of Tetrastigma hemsleyanum total extraction (TH-t) on the function of human γδT cells. Tetrastigma hemsleyanum total extraction (TH-t) was extracted from dried roots of Tetrastigma hemsleyanum with 80% ethanol. Human venous blood mononuclear cells (PBMC) were isolated using Ficoll separation solution. PBMCs were in orientational culture of γδT cells with the isopentenyl pyrophosphate method. Flow cytometry was used to detect the surface labeling rate of γδT CR before and after PBMC culture. And the percentage of CD107a, Granzyme B and Perforin on the surface of γδT cells was detected by fluorescently labeled monoclonal antibody. The effect of γδT cells induced by TH-t on the killing activity of tumor cell lines was examined by lactate dehydrogenase release assay. The CCK8 method was used to detect the effect of TH-t on the proliferation of γδT cells. MTT assay was used to detect the inhibition rate of TH-t on liver cancer (HepG2) cell line, gastric cancer (SGC-7901) cell line and breast cancer (MCF-7) cell line. The expression rates of γδTCR before PBMC culture were 3.12% and 90.46% after 10 days of directional culture. In the experiment of the effect of TH-t on the proliferation of γδT cells, TH-t at a concentration of 0.62 μg/mL induced the most obvious proliferation of γδT cells (44.50%) after 72 h of induction, which was significantly higher than that of the control group (3.50%) (P<0.05). When the TH-t concentration was 0.15 μg/mL, the positive expression rates of Perforin and granzymeB on the surface of induced γδT cells reached the highest values (76.90%±2.30% and 30.50%±1.30%, respectively), which was significantly higher than that of the control group was 65.40%±1.29% and 25.10%±2.30%, and there was a statistically significant difference between the groups (P<0.05). After induction by TH-t at a concentration of 0.125μg/mL, γδT cells had the highest cytotoxic activity against tumor HepG2, SGC-7901 and MCF-7 cells (72.10%, 52.30% and 79.10%, respectively), which was significantly higher than the control group (38.50%, 30.50% and 41.20%, respectively), there was a statistically significant difference between the groups (P<0.05). When the concentration of Th-t was≥9.75 μg/mL, it inhibited the three tumor cells. TH-t can promote the proliferation of γδT cells and increase the activity of killing tumor cells. A certain concentration of TH-t can inhibit the growth of HepG2, SGC-7901 and MCF-7 cells.
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