ÌÇ»ù»¯ÊÇÔÚøµÄ¿ØÖÆÏ£¬µ°°×Öʸ½¼ÓÉÏÌÇÀàµÄ¹ý³Ì£¬ÆðʼÓÚÄÚÖÊÍø£¬½áÊøÓڸ߶û»ùÌå¡£ ÔÚÌÇ»ùתÒÆø×÷ÓÃϽ«ÌÇתÒÆÖÁµ°°×ÖÊ£¬ºÍµ°°×ÖÊÉϵݱ»ùËá²Ð»ùÐγÉÌÇÜÕ¼ü¡£ ÕæºËÉúÎïÖУ¬Ò»°ëÒÔÉϵĵ°°×ÖÊ¿É·¢ÉúÌÇ»ù»¯ÐγÉÌǵ°°×£¬ÆäÖÐ90£¥ÊÇN-Ìǵ°°×¡£ÌÇ»ù»¯¾ßÓзdz£ÖØÒªºÍ¹ã·ºµÄ¹¦ÄÜ£º¢Ù ¶Ôϸ°û¾ßÓб£»¤¡¢Îȶ¨¼°ÆÁÕϵȶ෽Ãæ×÷Óã»¢Ú ¿É×÷ΪÍâÔ´ÐÔÊÜÌåµÄÌØÒìÐÔÅäÌ壬ijЩÌÇÁ´¿É×÷Ϊ¸÷ÖÖ²¡¶¾¡¢Ï¸¾ú¼°¼ÄÉúÎïµÄÌØÒìÊÜÌ壻¢Û ÌÇÁ´Ò²¿É×÷ΪÄÚÔ´ÐÔÊÜÌåµÄÌØÒìÐÔÅäÌ壬½éµ¼Çå³ý¡¢ÖÜת¼°°ûÄÚ´©ÐУ»¢Ü  ÌÇÁ´ÔÚÊܾ«¹ý³ÌÖÐÒ²Æð×ÅÖØÒªµÄ×÷Óá£

 

 

 

       Ò»°ãÇé¿öÏ£¬½«ÌÇ»ù»¯ÐÞÊηÖΪN- ÌÇ»ù»¯ºÍ O-ÌÇ»ù»¯Á½ÖÖ´óÀࣺN- ÌÇ»ù»¯ÊÇÖ¸ÌÇÁ´Óëµ°°×ÖÊÌ춬°±ËáÉϵݱ»ù»ùÍÅͨ¹ý¹²¼ÛÁ¬½Ó¶øÐγɵÄÌǵ°°×¡£Âú×ã°±»ùËáÐòÁÐAsn-X-Thr/Ser/Cys£¨X≠Pro£©£¬¾ßÓо­µäµÄºËÐÄÎåÌǽṹ£¨Man3GlcNAc2£©¡£O-ÌÇ»ù»¯£ºÍ¨¹ýÌÇÜÕ¼üÓëSer¡¢Thr¡¢ôÇ»ùÀµ°±Ëᣨ½ºÔ­£©»òTyr£¨ÌÇÔ­µ°°×£©µÄôÇ»ùÏàÁ¬¡£½Ï¶Ì£¬ÎÞºËÐĽṹ¡£Òò´Ë£¬O-ÌÇ»ù»¯µÄÌÇÁ´½á¹¹µÄ½âÎöÄѶÈÒª¸ßÓÚN- ÌÇ»ù»¯¡£Í¬Ê±£¬ÌÇ»ù»¯ÐÞÊξßÓкê¹ÛºÍ΢¹Û²»¾ùÒ»ÐÔ£¬ÏàͬµÄÌÇÁ´¿ÉÒÔÐÞÊÎͬһµ°°×µÄ²»Í¬Î»µã£¬Í¬Ò»Î»µãÒ²¿ÉÒÔÐÞÊβ»Í¬½á¹¹µÄÌÇÁ´£¬Æä½âÎöÄѶȷdz£´ó£¬³ÉΪÌÇ»ù»¯Ñо¿µÄÄѵãÎÊÌâ¡£

|   ÌØÉ«ÌÇ»ù»¯µ°°××éѧ

        ´«Í³µÄÌǵ°°×Ñо¿Æ½Ì¨£¬Ò»°ãͨ¹ýÌÇÜÕø½«¾ÛÌÇ´Ó¶àëÄÉÏÇгý£¨ÒªÃ´±£Áô¶àëÄ£¬ÒªÃ´±£ÁôÌÇÁ´£©£¬È»ºóͨ¹ýÖÊÆ×½âÎö»ñµÃ¶àëÄÉϵÄÌÇ»ù»¯ÐÞÊÎλµã£¬»òÕß½âÎöÌÇÁ´½á¹¹¡£ÕâЩ²ßÂÔҪôɥʧÁËÌǽṹÐÅÏ¢£¬ÒªÃ´¶ªÊ§Á˶àëÄλµãÐÅÏ¢£¬ÎÞ·¨×öµ½ÌǽṹÓë¶àëÄλµãµÄ¶ÔÓ¦·ÖÎö£¬Ê§È¥ÁËÌÇ»ù»¯µ°°××éѧÑо¿µÄºËÐÄÐÅÏ¢¡£

        k8¿­·¢(Öйú)ÀûÓÃøµÄÌØÒìÐÔ£¬½«µ°°×ÇгɶàëÄ£¬²¢Í¨¹ý¸»¼¯·½·¨»ñÈ¡º¬ÓоÛÌǵÄÌÇëÄ£¬ÀûÓøßÄÜÅöײËéÁÑÖÊÆ×½«ÌÇëÄÖð¼¶ËéÁÑ£¬»ñÈ¡ÖÊÆ׸÷¼¶Àë×ÓÐÅÏ¢¡£²¢Í¨¹ýרÀûÖÊÆ×Êý¾Ý±È¶ÔÈí¼þºÍ×Ô½¨µÄ70000ÌõÌÇëÄÊý¾Ý¿â£¬¶Ô¶àëĵÄÌÇ»ù»¯Î»µãºÍÌÇÁ´½á¹¹½øÐжԱȷÖÎö£¬¼È¿ÉÒÔ±£ÁôÌÇ»ù»¯Î»µãÐÅÏ¢£¬ÓÖ¿ÉÒÔ½âÎöÌǵĽṹ£¬ÔÚÌÇ»ù»¯¶Ô±È×éѧÑо¿ÖоßÓжÀÌصÄÓÅÊÆ¡£

       Í¬Ê±£¬k8¿­·¢(Öйú)»¹¿Éͨ¹ý¸ßͨÁ¿Äý¼¯ËØоƬÓëÌÇ»ù»¯µ°°××éÖÊÆ×¼¼ÊõµÄ×éºÏ£¬Íê³É¸üΪ¸´ÔÓµÄÑо¿¿ÎÌâ¡£ÎÒÃÇ¿ÉÒÔͨ¹ý¸ßͨÁ¿Äý¼¯ËØоƬ¼¼Êõ¶Ô´óÅúÁ¿ÁÙ´²Ñù±¾ÖеÄÌǵ°°×Æ×½øÐвîÒìɸÑ¡£¬ÕÒµ½ºòÑ¡±êÖ¾ÎÔÙ¶ÔÌض¨Äý¼¯ËØʶ±ðµÄÌǵ°°×½øÐÐÌÇ»ù»¯µ°°××éÖÊÆ×½âÎö»ñµÃÍêÕûµÄÌÇÆ׽ṹºÍÌÇ»ù»¯Î»µãͼÆס£

|   Êý¾Ý½á¹û

1.  ʵÑé²½Ö裨ÖÐÓ¢ÎÄ£©

2.  ÖÊÆ×ÒDzɼ¯µÄԭʼÊý¾Ý

3.  »ùÓÚ°ÐÏò-ÓÕ¶ü¿âËÑË÷ºÍÆ×ͼˮƽFDR¿ØÖƵÄÍêÕûN-ÌÇëÄIDsµÄÈ«Ãæ¼ø¶¨ÐÅÏ¢£¬°üÀ¨¶àëĹǼܣ¨°±»ùËáÐòÁС¢Ç±ÔÚN-ÌÇ»ù»¯Î»µã£©ºÍN-Á¬½ÓÌDz¿·Ö£¨µ¥ÌÇ×é³É¡¢Ç±ÔÚÐòÁкÍÌÇÜÕ¼üÁ¬½Ó½á¹¹£©£»ÆäÖÐN-ÌÇ»ù»¯Î»µã½øÒ»²½µÃµ½Î»µã¾ö¶¨ÐÔËéƬÀë×ÓÈ·ÈÏ£¬ÐòÁкÍÁ¬½Ó½á¹¹½øÒ»²½µÃµ½½á¹¹Õï¶ÏËéƬÀë×ÓµÄÈ·ÈÏ¡£

4.  ²îÒìµ°°×ÉúÐÅ·ÖÎö½á¹û

|   Ñù±¾ÒªÇó

Ñù±¾ÐÎʽ£ºÐÂÏÊÀ䶳×éÖ¯£¬FFPE×éÖ¯£¬ÌåÒº£¨Ñª½¬ÑªÇå¡¢ÄòÒº£©£¬Ï¸°û£¬ÍêÕûµ°°×£¬¶àëÄ

Ñù±¾Á¿£º¶¯Îï×éÖ¯>1 g£»Ö²Îï×éÖ¯>200 mg£»ÑªÒºÑù±¾>1 mL£¨×¢ÒâÓÃEDTA·ÀÄý£©£»ÑªÇå>0.5 mL£»ÄòÒº>2 mL£»Ï¸°û>5×10^7£»½Íĸ/΢ÉúÎï>200 mg

×¢ÒâÒªµã£º¾¡Á¿±ÜÃâʹÓñíÃæ»îÐÔ¼Á£¨SDS¡¢Triton-X£©ºÍÎÞ»úÑÎ

|   ¾­µä°¸Àý

      ´ÓÁÙ´²Ñù±¾ÖзÖÎö·¢ÏÖ£¬ÈéÏÙ°©ÔÚ·¢Éú·¢Õ¹¹ý³ÌÖдæÔÚÒ»ÖÖ½Ð×÷ƽ·ÖÐÍGlcNAcµÄN-ÌÇ»ù»¯ÐÞÊεÄˮƽÏÔÖøϽµµÄÏÖÏ󣬲¢ÓëÈéÏÙ°©Éú´æÆÚÃÜÇÐÏà¹Ø¡£½øÒ»²½Ñо¿·¢ÏÖ£¬ÇÖÏ®ÄÜÁ¦½ÏÇ¿µÄÈéÏÙ°©Ï¸°û·ÖÃÚµÄϸ°ûÍâÄÒÅÝÉϵÄĤµ°°×Ò²µÍ±í´ïƽ·ÖÐÍGlcNAcÐÞÊΡ£ÓÉ´ËÌá³ö¼ÙÉ裬¿ÉÄÜתÒÆÄÜÁ¦½ÏÈõµÄÈéÏÙ°©Ï¸°û£¨ÊÜÌåϸ°û£©µÄתÒÆÄÜÁ¦»á±»×ªÒÆÄÜÁ¦½ÏÇ¿µÄÈéÏÙ°©Ï¸°û£¨¹©Ìåϸ°û£©Í¨¹ý·ÖÃÚÍâÃÚÌåµÄ·½Ê½¼¤»î¡£
 

       ÓÚÊÇ£¬Ñо¿ÈËÔ±ÀûÓÃÌض¨Äý¼¯ËØLectin¸»¼¯½áºÏÖÊÆ×·ÖÎöµÄ²ßÂÔ·ÖÎöÁËÍâÃÚÌå´¦ÀíºóµÄÈéÏÙ°©Ï¸°û±íÃæÌǵ°°×µÄ¸Ä±ä£¬·¢ÏÖϸ°ûĤµ°°×integrin β1¿ÉÄÜÊܵ½Æ½·ÖÐÍGlcNAcÐÞÊεĵ÷¿Ø¡£½øÒ»²½£¬Í¨¹ýÌÇ»ù»¯µ°°××éÖÊÆ×·ÖÎö£¬½âÎöÁËGlcNAcÐÞÊεÄÌǽṹ£¬Ö¤ÊµÁËÆäÓëintegrin β1½áºÏµÄÄÜÁ¦¼°½áºÏλµã¡£Í¨¹ý×è¶ÏʵÑ飬µÃ³ö×îÖÕ½áÂÛ£ºÆ½·ÖÐÍGlcNAcÐÞÊξßÓÐͨ¹ýÒÖÖÆÌÇÁ´µÄÑÓÉ죬¸Ä±äµ°°×ÖʵÄÌÇ»ù»¯ÐÞÊÎˮƽ£¬´Ó¶øÓ°ÏìÊÜÌåϸ°ûĤµ°°×integrin β1¼°ÆäÏÂÓÎFAK-AKTͨ·µÄ¼¤»î£¬½ø¶øÒÖÖÆÊÜÌåÈéÏÙ°©Ï¸°ûÇÖÏ®µÄÄÜÁ¦¡£È»¶ø£¬×ªÒÆÄÜÁ¦½ÏÇ¿µÄÈéÏÙ°©Ï¸°û¾ùµÍ±í´ïƽ·ÖÐÍGlcNAcÐÞÊΣ¬Ê¹µÃÆä·ÖÃÚµÄÍâÃÚÌå¾ß±¸ÁËÓÕµ¼ÊÜÌåÈéÏÙ°©Ï¸°û¼ÓËÙתÒƵÄÄÜÁ¦¡£

|   Ïà¹ØÎÄÏ×

[1]. Xu Junying,Y ang Xuejing, Mao Yong, et al. Removal of N-Linked Glycosylation Enhances PD-L1 Detection in Colon Cancer: Validation Research Based on Immunohistochemistry Analysis. Technol Cancer Res Treat, 2021, 20: 15330338211019442.

[2]. Tan Zengqi, Cao Lin, Wu Yurong, et al. Bisecting GlcNAc modification diminishes the pro-metastatic functions of small extracellular vesicles from breast cancer cells. J Extracell Vesicles, 2020, 10: e12005.

[3]. Jingyun Yang, Wei Wang, Zimin Chen, et al. A vaccine targeting the RBD of the S protein of SARS-CoV-2 induces protective immunity. Nature, 2020, 586: 572–577.

[4]. Wang Yue, Xu Feifei, Chen Yun, et al. A quantitative N-glycoproteomics study of cell-surface N-glycoprotein markers of MCF-7/ADR cancer stem cells.[J] .Anal Bioanal Chem, 2020, 412: 2423-2432.

[5]. Xu Feifei, Wang Yue, Xiao Kaijie, et al. Quantitative site- and structure-specific N-glycoproteomics characterization of differential N-glycosylation in MCF-7/ADR cancer stem cells. Clin Proteomics, 2020, 17: 3.

[6]. Xue Bingbing, Xiao Kaijie, Wang Yue, et al. Site- and structure-specific quantitative N-glycoproteomics study of differential N-glycosylation in MCF-7 cancer cells. J Proteomics, 2020, 212: 103594.