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一、浙江大學附屬第二醫(yī)院乳腺外科；浙江大學附屬第二醫(yī)院浙江省療法腫瘤微環(huán)境與免疫重點實驗室；浙江省人民醫(yī)院，浙江省人民醫(yī)院，杭州醫(yī)學院附屬人民醫(yī)院，浙江省個體化醫(yī)學腫瘤分子診斷與診斷重點實驗室；紹興大學附屬醫(yī)院甲狀腺乳腺外科在2020-2-19聯合發(fā)表標題為Breast cancer-derived exosomes transmit lncRNA SNHG16 to induce CD73+γδ1 Treg cells（乳腺癌衍生的外泌體傳遞lncRNA SNHG16誘導CD73 +γδ1 Treg細胞）的文章到nature /sigtrans，文章已被接受；

二、本研究使用中使用Invigentech（英克）公司INVI DNA RNA 轉染試劑轉染質粒DNA、siRNA到Vδ1 T 免疫T細胞里面；

三、本文中轉染質粒DNA、siRNA，轉染細胞數量信息如下：

A. 將全長2435 bp的序列克隆到pCR3.1載體中構建SNHG16過表達載體;

B. siRNA序列：SNHG16-homo-349，5′GCCUCUGCUGCUAAUUGUUTT-3'; SNHG16-homo-867， 5′-CCAAGGAGGGACUGUUUAATT-3′和SNHG16-homo-2004，5′-CCCAGUGUUGACUCACCAATT-3；

C. 轉染細胞用量：6孔板里面1 × 106 cells/well；

四、發(fā)表文章部分內容如下：

Breast cancer-derived exosomes transmit lncRNA SNHG16 to induce CD73+γδ1 Treg cells

Plasmid construction and siRNA silencing A full-length 2435 bp sequence was cloned into the pCR3.1 vector to

construct the SNHG16 overexpression vector. The sequences of SNHG16-specifific siRNAs were as follows: SNHG16-homo-349, 5′GCCUCUGCUGCUAAUUGUUTT-3′; SNHG16-homo-867, 5′-CCAAGGAGGGACUGUUUAATT-3′ and SNHG16-homo-2004, 5′-CCCAGUGUUGACUCACCAATT-3′. The miR-16–5p mimics, inhibitor and negative

controls were purchased from GenePharma (SupplementaryTable S3).

To manipulate the expression of miR-16–5p in Vδ1 T cells, Vδ1T cells were sorted from peripheral blood and seeded into six-wellplates at 1 × 106 cells/well with 1 ml of medium supplementedwith 10% FBS, 10 μg/ml CD3, 10 μg/ml CD28 and 40 U/mL IL-2.Then, transfection reagent (INVI DNA RNA Transfection Reagent,Invigentech, USA) and miR-16–5p mimics/NC/inhibitor/inhibitor NC (20 μm) were incubated with the cells for 48 h, after which the cells were collected for further experiments.

but not Vδ2+ T cells were revealed to comprise the majority of TILs in all three BC molecular subtypes when gated on CD3 (p < 0.01, Fig. 1b, c). In addition, immunoflfluorescence of frozen sections further confifirmed the dominant distribution of Vδ1+T cells in BC (Fig. 1d). 

To further identify the function of breast cancer-infifiltrating γδ1T cells, BC cell lines (MCF-7, T-47D, MDA-MB-231 and MDA-MB-468) were co-cultured with freshly isolated Vδ1+ T cells frombreast tumour tissue (E:T ratios: 1:1, 5:1 and 10:1). Upon plotting the cell growth curve, the data show that γδ1 T cells did not exert

invigentech（英克）INVI DNA RNA轉染試劑信息如下