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\u7684\u7d27\u5bc6\u94fa\u8def\u77f3\u6392\u5217\uff0c\u6838\u6d46\u6bd4\u5927\u4e8e MIA-PACA-2\uff08\u8001\u5e74\uff09\u3001\u5c0f\u4e8e JF-305\uff08\u4e2d\u5e74\u901a\u7528\u578b\uff09\uff0c\u4f53\u73b0\u80f0\u7ba1\u764c\u7ec6\u80de\u805a\u56e2\u751f\u957f\u7684\u5f62\u6001\u7279\u5f81\uff1b\u4e0e MIA-PACA-2\uff08\u534a\u8d34\u58c1 \/ \u677e\u6563\u805a\u56e2\uff09\u76f8\u6bd4\uff0c\u65e0\u60ac\u6d6e\u7ec6\u80de\u4e14\u805a\u56e2\u66f4\u7d27\u5bc6\uff0c\u4e0e JF-305\uff08\u8d34\u58c1 \/ \u65e0\u805a\u56e2\uff09\u76f8\u6bd4\uff0c\u805a\u56e2\u751f\u957f\u7279\u5f81\u663e\u8457\uff1b\u53ef\u901a\u8fc7 \u201c\u4e0a\u76ae\u6837\u7d27\u5bc6\u805a\u56e2 + KRAS p.Gly12Asp \u7a81\u53d8 + \u80f0\u7ba1\u6807\u5fd7\u7269\uff08\u5982 CK19\uff09\u9633\u6027\u201d \u5feb\u901f\u9274\u522b\uff0c\u540c\u65f6\u4e0e MIA-PACA-2 \u901a\u8fc7\u751f\u957f\u7279\u6027\uff08\u7eaf\u8d34\u58c1 vs \u534a\u8d34\u58c1\uff09\u3001\u4e0e JF-305 \u901a\u8fc7\u805a\u56e2\u7279\u6027\uff08\u6709 vs \u65e0\uff09\u8fdb\u4e00\u6b65\u533a\u5206\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u4ee3\u6570\uff1a10 \u4ee3\u4ee5\u5185\uff08\u4f4e\u4ee3\u6570\u7ec6\u80de\u80fd\u66f4\u597d\u4fdd\u7559 \u201c\u80f0\u7ba1\u7279\u6027\u3001\u805a\u56e2\u751f\u957f\u3001KRAS \u7a81\u53d8\u72b6\u6001\u201d \u6838\u5fc3\u5c5e\u6027\uff0c\u5efa\u8bae\u4f20\u4ee3\u6b21\u6570\u4e25\u683c\u63a7\u5236\u5728 10 \u4ee3\u4ee5\u5185\uff0c\u9632\u6b62\u957f\u671f\u4f20\u4ee3\u5bfc\u81f4\u7279\u6027\u6f02\u79fb\uff0c\u5982\u805a\u56e2\u80fd\u529b\u51cf\u5f31\uff08\u805a\u56e2\u76f4\u5f84\uff1c30\u03bcm\uff09\u3001KRAS \u7a81\u53d8\u4e30\u5ea6\u4e0b\u964d\uff08\u4f4e\u4e8e\u521d\u59cb\u4ee3\u6b21 80%\uff09\u3001\u500d\u589e\u65f6\u95f4\u7f29\u77ed\uff08\uff1c35 \u5c0f\u65f6\uff09\uff0c\u786e\u4fdd\u7ec6\u80de\u59cb\u7ec8\u8d34\u5408\u80f0\u7ba1\u764c\u53ca KRAS \u7814\u7a76\u9700\u6c42\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u80cc\u666f\u4ecb\u7ecd\uff1aPANC-1 \u7ec6\u80de\u6e90\u81ea 56 \u5c81\u767d\u4eba\u7537\u6027\u80f0\u7ba1\u4e0a\u76ae\u7ec6\u80de\u764c\uff0c\u6838\u5fc3\u7279\u6027\u53ca\u5e94\u7528\u5982\u4e0b\uff1a\u2460\u75c5\u7406\u4e0e\u5206\u5b50\u7279\u8272\uff1a\u4f5c\u4e3a\u80f0\u7ba1\u764c\u7ecf\u5178\u6a21\u578b\uff0c\u542b\u80f0\u817a\u764c\u9ad8\u9891 KRAS p.Gly12Asp \u70ed\u70b9\u7a81\u53d8\uff08\u7ea6 90% \u80f0\u7ba1\u764c\u5b58\u5728\u8be5\u7a81\u53d8\uff09\uff0cG6PD \u4f4e\u6cf3\u52a8\u6027 B \u578b\uff08\u9176\u6d3b\u6027\u7279\u5f81\u53ef\u8f85\u52a9\u8eab\u4efd\u9a8c\u8bc1\uff09\uff0c\u662f\u7814\u7a76 KRAS \u9a71\u52a8\u80bf\u7624\u7684\u7406\u60f3\u5de5\u5177\uff1b\u2461\u751f\u957f\u7279\u6027\u4ef7\u503c\uff1a\u6613\u805a\u56e2\u751f\u957f\u6a21\u62df\u80f0\u7ba1\u764c\u4e34\u5e8a\u75c5\u7406\u4e2d\u7684\u817a\u7ba1\u6837\u7ed3\u6784\uff0c\u53ef\u7528\u4e8e\u63a2\u7d22\u7ec6\u80de\u95f4\u8fde\u63a5\uff08\u5982 E-cadherin\uff09\u4e0e\u805a\u56e2\u7684\u5173\u8054\uff1b\u2462\u5e94\u7528\u573a\u666f\uff1a\u5e7f\u6cdb\u7528\u4e8e\u80f0\u7ba1\u764c\u57fa\u7840\u673a\u5236\uff08\u5982 KRAS-RAF-MEK \u901a\u8def\uff09\u3001KRAS \u6291\u5236\u5242\u7b5b\u9009\u3001\u805a\u56e2\u751f\u957f\u8c03\u63a7\u7814\u7a76\uff0c\u540c\u65f6\u53ef\u4f5c\u4e3a \u201c\u80f0\u7ba1\u764c\u6807\u51c6\u7ec6\u80de\u7cfb\u201d \u7528\u4e8e\u591a\u4e2d\u5fc3\u5b9e\u9a8c\u7ed3\u679c\u6bd4\u5bf9\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u91cd\u8981\u63d0\u9192\uff08\u6838\u5fc3\u64cd\u4f5c\u8b66\u793a\uff0c\u4e0e MIA-PACA-2 \u5bf9\u6bd4\uff09\uff1a\u2460\u6d88\u5316\u64cd\u4f5c\uff1a\u56e0\u6613\u805a\u56e2\uff0c\u9700\u4f7f\u7528 0.25% \u80f0\u9176 &#8211; EDTA \u6d88\u5316\u6db2\uff08\u53ef\u6dfb\u52a0 0.01% DNase I \u9632\u6b62\u7ec6\u80de\u56e2\u7c98\u8fde\uff09\uff0c37\u2103\u5b75\u80b2 4-5 \u5206\u949f\uff0c\u671f\u95f4\u6bcf 2 \u5206\u949f\u8f7b\u67d4\u62cd\u6253\u57f9\u517b\u74f6\u58c1 1 \u6b21\uff0c\u955c\u4e0b\u89c2\u5bdf\u7ec6\u80de\u56e2\u5206\u6563\u4e3a\u5355\u4e2a\u7ec6\u80de\u540e\u7acb\u5373\u7ec8\u6b62\uff1b\u7ec8\u6b62\u540e\u7528\u542b 10% FBS \u7684 DMEM \u57f9\u517b\u57fa\u4e2d\u548c\u80f0\u9176\uff0c\u8f7b\u8f7b\u5439\u6253 3-5 \u6b21\uff08\u907f\u514d\u8fc7\u5ea6\u5439\u6253\u5bfc\u81f4\u7ec6\u80de\u635f\u4f24\uff09\uff0c\u786e\u4fdd\u5355\u7ec6\u80de\u60ac\u6db2\u6bd4\u4f8b\u226590%\uff1b\u2461\u94fa\u677f\u64cd\u4f5c\uff1a\u7ec6\u80de\u8ba1\u6570\u540e\uff0c\u6309\u6240\u9700\u5bc6\u5ea6\u7a00\u91ca\u4e3a\u5355\u7ec6\u80de\u60ac\u6db2\uff0c\u94fa\u677f\u65f6\u8f7b\u8f7b\u6447\u6643\u57f9\u517b\u74f6\uff0c\u4f7f\u7ec6\u80de\u5747\u5300\u5206\u5e03\uff0c\u907f\u514d\u805a\u56e2\u6c89\u6dc0\u5728\u74f6\u5e95\u5c40\u90e8\uff1b\u94fa\u677f\u540e 24 \u5c0f\u65f6\u5185\u4e0d\u79fb\u52a8\u57f9\u517b\u74f6\uff0c\u9632\u6b62\u7ec6\u80de\u805a\u56e2\u4f4d\u7f6e\u6539\u53d8\uff1b\u2462\u805a\u56e2\u76d1\u6d4b\uff1a\u4f20\u4ee3\u540e 48 \u5c0f\u65f6\u955c\u68c0\uff0c\u82e5\u4ecd\u51fa\u73b0\u5927\u91cf\u5927\u805a\u56e2\uff08\u76f4\u5f84\uff1e150\u03bcm\uff09\uff0c\u9700\u91cd\u65b0\u4f18\u5316\u6d88\u5316\u65f6\u95f4\uff08\u5ef6\u957f 1-2 \u5206\u949f\uff09\u6216\u6dfb\u52a0 DNase I\uff0c\u786e\u4fdd\u540e\u7eed\u5b9e\u9a8c\uff08\u5982\u836f\u7269\u7b5b\u9009\u3001\u8f6c\u67d3\uff09\u7684\u7ec6\u80de\u5747\u4e00\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">STR \u4f4d\u70b9\u4fe1\u606f\uff1aAmelogenin\uff1aX\uff08\u7ed3\u5408\u6027\u522b\u6807\u6ce8 \u201c\u7537\u201d\uff0c\u5b58\u5728 Y \u67d3\u8272\u4f53\u7f3a\u5931\u53ef\u80fd\uff0c\u5efa\u8bae\u901a\u8fc7 SRY \u57fa\u56e0 PCR \u9a8c\u8bc1\u6027\u522b\uff0c\u907f\u514d\u6838\u578b\u8bef\u5224\uff09; CSF1PO\uff1a10\uff0c12\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD13S317\uff1a11\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD16S539\uff1a11\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD18S51\uff1a12\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD19S433\uff1a11\uff0c16\uff08\u7f55\u89c1\u7b49\u4f4d\u57fa\u56e0\uff0c\u6838\u5fc3\u9274\u522b\u4f4d\u70b9\uff09\uff1bD21S11\uff1a28\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD2S1338\uff1a23\uff0c24\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD3S1358\uff1a17\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD5S818\uff1a11\uff0c13\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD7S820\uff1a8\uff0c10\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bD8S1179\uff1a14\uff0c15\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bFGA\uff1a21\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bTH01\uff1a7\uff0c8\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bTPOX\uff1a8\uff0c11\uff08\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1bvWA\uff1a15\uff08\u7eaf\u5408\u5b50\uff0c\u4e0e MIA-PACA-2 \u7684\u672a\u660e\u786e\u4f4d\u70b9\u5dee\u5f02\uff09\uff1b\uff08STR \u4f4d\u70b9\u542b\u7f55\u89c1\u7b49\u4f4d\u57fa\u56e0\uff08D19S433\uff1a16\uff09\uff0c\u4e0e MIA-PACA-2\u3001JF-305 \u7684 STR \u56fe\u8c31\u5dee\u5f02\u663e\u8457\uff0c\u53ef\u901a\u8fc7\u4e0e ATCC CRL-1469 \u6570\u636e\u6bd4\u5bf9\uff0c\u786e\u8ba4\u7ec6\u80de\u8eab\u4efd\u53ca\u80f0\u7ba1\u764c\u5c5e\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57fa\u56e0\u53d8\u5f02\u4fe1\u606f\uff1ap.Gly12Asp (c.35G&gt;A); ClinVar=VCV000012582; Zygosity=Heterozygous\uff08KRAS \u70ed\u70b9\u7a81\u53d8\uff09\uff1a\u2460\u7a81\u53d8\u7279\u5f81\uff1aKRAS \u57fa\u56e0\u7b2c 12 \u4f4d\u7518\u6c28\u9178\uff08Gly\uff09\u7a81\u53d8\u4e3a\u5929\u51ac\u6c28\u9178\uff08Asp\uff09\uff0c\u4e3a\u80f0\u817a\u764c\u9ad8\u9891\u9a71\u52a8\u7a81\u53d8\uff08\u6742\u5408\u5b50\uff0c\u4fdd\u7559\u91ce\u751f\u578b allele\uff09\uff0c\u53ef\u901a\u8fc7 Sanger \u6d4b\u5e8f\u6216\u6570\u5b57 PCR \u9a8c\u8bc1\u7a81\u53d8\u4e30\u5ea6\uff08\u5efa\u8bae\u6bcf 5 \u4ee3\u68c0\u6d4b 1 \u6b21\uff09\uff1b\u2461\u529f\u80fd\u5f71\u54cd\uff1a\u6fc0\u6d3b KRAS-RAF-MEK-ERK \u4fe1\u53f7\u901a\u8def\uff0c\u4fc3\u8fdb\u7ec6\u80de\u589e\u6b96\u3001\u6297\u51cb\u4ea1\u53ca\u805a\u56e2\u751f\u957f\uff0c\u662f\u80f0\u7ba1\u764c\u6076\u6027\u8fdb\u5c55\u7684\u5173\u952e\u9a71\u52a8\u56e0\u7d20\uff1b\u2462\u5e94\u7528\u573a\u666f\uff1aa. \u9776\u5411\u6cbb\u7597\u5b9e\u9a8c\uff1a\u4f7f\u7528 KRAS G12D \u6291\u5236\u5242\uff08\u5982 MRTX1133\uff09\u5904\u7406\u7ec6\u80de\uff0c\u68c0\u6d4b\u589e\u6b96\u6291\u5236\u7387\uff08CCK-8\uff09\u53ca\u901a\u8def\u6d3b\u6027\uff08p-ERK Western Blot\uff09\uff0c\u8bc4\u4f30\u836f\u7269\u7597\u6548\uff1bb. \u673a\u5236\u7814\u7a76\uff1a\u901a\u8fc7 siRNA \u5e72\u6270 KRAS \u8868\u8fbe\uff0c\u63a2\u7d22\u7a81\u53d8\u578b KRAS \u5bf9\u80f0\u7ba1\u764c\u805a\u56e2\u751f\u957f\u3001\u81f4\u7624\u6027\u7684\u8c03\u63a7\u4f5c\u7528\uff1b\u2463\u6ce8\u610f\u4e8b\u9879\uff1a\u957f\u671f\u57f9\u517b\u9700\u76d1\u6d4b\u7a81\u53d8\u4e30\u5ea6\uff0c\u907f\u514d\u91ce\u751f\u578b\u7ec6\u80de\u514b\u9686\u4f18\u52bf\u751f\u957f\u5bfc\u81f4\u7a81\u53d8\u4e30\u5ea6\u4e0b\u964d\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u67d3\u8272\u4f53\uff1a60~70\uff08\u5f02\u500d\u4f53\uff0c\u67d3\u8272\u4f53\u6570\u76ee\u4ecb\u4e8e MIA-PACA-2 \u7684\u672a\u660e\u786e\u6838\u578b\u4e0e JF-305 \u7684\u672a\u660e\u786e\u6838\u578b\u4e4b\u95f4\uff09\uff1a\u2460\u6838\u578b\u7279\u5f81\uff1a\u67d3\u8272\u4f53\u6a21\u5f0f\u6570\u76ee 63\uff08\u542b 3 \u4e2a\u72ec\u7279\u6807\u8bb0\u67d3\u8272\u4f53 + 1 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