CAJ | 학술논문

HeLa细胞KB株、X株、NM20／X株、H株的染色体众数依次为60±3（超二倍体）、62±3（超二倍体）、68±3（超二倍体和亚四倍体）和78±2（亚四倍体），所占比率分别为72%～76%，69%，52%和40%。在纯化3代的肿瘤阴性对照二倍体猫肾（染色体众数38所占比率80%）和犬肾原代细胞皮下接种裸鼠的致癌／致瘤率分别为0%（0／22）和0%（0／10），X株HeLa细胞冻融裂解物皮下接种裸鼠产生进行性缩小肿瘤的比率为20%（1／5）的前提下，HeLa细胞KB株、X株、NM20／X株皮下接种裸鼠产生进行性生长恶性肿瘤的比率分别为100%（10／ 10），100%（25／25）和100%（5／5），H株细胞皮下接种裸鼠产生恶性肿瘤的比率为50%（5／10）。其中，只有HeLa细胞KB株10～11代（染色体结构畸变率高达20%，出现18%双着丝点和2%断片）以超高数量接种的1组4只裸鼠（0.17ml 12.75×107／鼠）才均形成MRT，特别是从染色体众数不同的3株HeLa细胞中筛选出致瘤性强的X株，连传20代后皮下接种裸鼠形成低分化肿瘤，定名为NM20／X株0代，其染色体众数因经裸鼠体内传代选育而明显增大（染色体众数为68±3，所占比率为52%），实体瘤手术切除后体外连传11代，再皮下接种裸鼠均形成MRT（5／5），但要求完形活细胞接种量要大（5～12×107／鼠），肿瘤产生迅速，生长快速，血管丰富，供血充分，接种细胞后16～22天肿瘤长径×短径均值基本达到30mm×20mm标准。这种MRT在模型动物体内的首次发现，说明宫颈上皮可能是MRT的起源组织之一，从而为弄清MRT（特别是子宫MRT）的起源问题提供了机会，开辟了组织发生清楚的细胞系用于MRT起源研究的新阶段。本研究结果表明，不同核型细胞的致瘤性不同，肿瘤细胞系高变异率株可在裸鼠体内快速选育成功，染色体结构畸变率特别高可能增加致癌性；HeLa细胞染色体遗传特征决定致瘤性质，细胞染色体数目变异大小和致癌／致瘤性强弱相关，致瘤性由弱到强的顺序依次为KB株，X株和NM20／X株，但H株有例外，尚待进一步实验确定。 HeLa細胞KB株、X株、NM20／X株、H株的染色體衆數依次為60±3（超二倍體）、62±3（超二倍體）、68±3（超二倍體和亞四倍體）和78±2（亞四倍體），所佔比率分彆為72%～76%，69%，52%和40%。在純化3代的腫瘤陰性對照二倍體貓腎（染色體衆數38所佔比率80%）和犬腎原代細胞皮下接種裸鼠的緻癌／緻瘤率分彆為0%（0／22）和0%（0／10），X株HeLa細胞凍融裂解物皮下接種裸鼠產生進行性縮小腫瘤的比率為20%（1／5）的前提下，HeLa細胞KB株、X株、NM20／X株皮下接種裸鼠產生進行性生長噁性腫瘤的比率分彆為100%（10／ 10），100%（25／25）和100%（5／5），H株細胞皮下接種裸鼠產生噁性腫瘤的比率為50%（5／10）。其中，隻有HeLa細胞KB株10～11代（染色體結構畸變率高達20%，齣現18%雙著絲點和2%斷片）以超高數量接種的1組4隻裸鼠（0.17ml 12.75×107／鼠）纔均形成MRT，特彆是從染色體衆數不同的3株HeLa細胞中篩選齣緻瘤性彊的X株，連傳20代後皮下接種裸鼠形成低分化腫瘤，定名為NM20／X株0代，其染色體衆數因經裸鼠體內傳代選育而明顯增大（染色體衆數為68±3，所佔比率為52%），實體瘤手術切除後體外連傳11代，再皮下接種裸鼠均形成MRT（5／5），但要求完形活細胞接種量要大（5～12×107／鼠），腫瘤產生迅速，生長快速，血管豐富，供血充分，接種細胞後16～22天腫瘤長徑×短徑均值基本達到30mm×20mm標準。這種MRT在模型動物體內的首次髮現，說明宮頸上皮可能是MRT的起源組織之一，從而為弄清MRT（特彆是子宮MRT）的起源問題提供瞭機會，開闢瞭組織髮生清楚的細胞繫用于MRT起源研究的新階段。本研究結果錶明，不同覈型細胞的緻瘤性不同，腫瘤細胞繫高變異率株可在裸鼠體內快速選育成功，染色體結構畸變率特彆高可能增加緻癌性；HeLa細胞染色體遺傳特徵決定緻瘤性質，細胞染色體數目變異大小和緻癌／緻瘤性彊弱相關，緻瘤性由弱到彊的順序依次為KB株，X株和NM20／X株，但H株有例外，尚待進一步實驗確定。
HeLa세포KB주、X주、NM20／X주、H주적염색체음수의차위60±3（초이배체）、62±3（초이배체）、68±3（초이배체화아사배체）화78±2（아사배체），소점비솔분별위72%～76%，69%，52%화40%。재순화3대적종류음성대조이배체묘신（염색체음수38소점비솔80%）화견신원대세포피하접충라서적치암／치류솔분별위0%（0／22）화0%（0／10），X주HeLa세포동융렬해물피하접충라서산생진행성축소종류적비솔위20%（1／5）적전제하，HeLa세포KB주、X주、NM20／X주피하접충라서산생진행성생장악성종류적비솔분별위100%（10／ 10），100%（25／25）화100%（5／5），H주세포피하접충라서산생악성종류적비솔위50%（5／10）。기중，지유HeLa세포KB주10～11대（염색체결구기변솔고체20%，출현18%쌍착사점화2%단편）이초고수량접충적1조4지라서（0.17ml 12.75×107／서）재균형성MRT，특별시종염색체음수불동적3주HeLa세포중사선출치류성강적X주，련전20대후피하접충라서형성저분화종류，정명위NM20／X주0대，기염색체음수인경라서체내전대선육이명현증대（염색체음수위68±3，소점비솔위52%），실체류수술절제후체외련전11대，재피하접충라서균형성MRT（5／5），단요구완형활세포접충량요대（5～12×107／서），종류산생신속，생장쾌속，혈관봉부，공혈충분，접충세포후16～22천종류장경×단경균치기본체도30mm×20mm표준。저충MRT재모형동물체내적수차발현，설명궁경상 피가능시MRT적기원조직지일，종이위롱청MRT（특별시자궁MRT）적기원문제제공료궤회，개벽료조직발생청초적세포계용우MRT기원연구적신계단。본연구결과표명，불동핵형세포적치류성불동，종류세포계고변이솔주가재라서체내쾌속선육성공，염색체결구기변솔특별고가능증가치암성；HeLa세포염색체유전특정결정치류성질，세포염색체수목변이대소화치암／치류성강약상관，치류성유약도강적순서의차위KB주，X주화NM20／X주，단H주유예외，상대진일보실험학정。
Under the prerequisite that the incidence of cancer or tumor in negatively?controllednudemiceinoculatedsubcutaneously with feline or canine kidney cell cultures purified in vitro at passage 3 or higher (the modal chromosome number of FKC on passage 3 was 38 of diploid at the rate of 80%) was 0%(0／22) and 0%(0／10) respectively, and the incidence of progressively negative growing tumor in controlled nude mice inoculated subcutaneously with repeatedly-frozen-andthawed-HeLa cell cultures of X strain was 20%(1／5), the negative growing malignant tumor (MT) was found in half of the nude mice inoculated subcutaneously with HeLa cell cultures of H strain(with modal chromosome number of 78±2 of sub-tetraploid at the rate of 40%), the progressively-growing malignant tumor was found in all the other 40 nude mice inoculated subcutaneously with HeLa cell cultures of other strains, with the incidence of MT in nude mice with KB strain (with modal chromosome number of 60±3 of hyperdiploid at the rate of 72%～76%) 10／10, the incidence of poorly-differentiated MT originated from epithelia in nude mice with X strain (with modal chromosomal number of 62±3 of hyperdiploid at the rate of 69%) 25／25, and the incidence of MRT in nude mice with in vitro cultured tumor cell NM20／X strain (with modal chromosome number of 68±3 of both hyperdiploid and subtetraploid at the rate of 52%) 5／5. After being continuously cultivated for 20 passages in vitro, HeLa cell of X strain was subcutaneously inoculated into nude mice and cultivated for 1 passage in vivo within 15 days, and then the developed growing MT was collected as HeLa cell of NM20／X strain on passage 0 and continuously cultivated for 11 passages to prepare for transplanting into nude mice again. Therefore,the highly variable strain of HeLa cells can be successfully selected by alternate cultivation in vitro and in vivo. Occasionally in another experiment, the progressivelyrowing MRT was found in all the 4 nude mice of one test group inoculated subcutaneously with 0.17ml cellultures of super杊igh density containing 12.75×107HeLa cells of KB strain on passages 10～11(with the rate of chromosome aberration high to 20% on passages 10～11 including 18% dicentric chromosome and 2% breakage chromosome). Although the incidence of MRT in nude mice inoculated subcutaneously with violently variable HeLa cells of NM20／X strain on passage 11, HeLa cells of KB strain on passages 10～11 reaches 100%(5／5) & 100%(4／4) respectively, yet it is requested that the inoculated live cell number is huge (5～12×107 cells per nude mouse), the tumor emerges immediately, develops violently, grows very fast, and has an extremely aggressive malignancy, the tumor is rich in the blood vessel giving a full supply of blood for it, and the mean value of major diameter X minor diameter of the tumor is essentially up to the standard of 30mm×20mm in 16～22 days after the inoculation of the cells into the nude mice. The first finding of MRT in model animals provides an opportunity for answering the origin problem of MRT. Based on this reason, human uterus vertical epithelium may be an original tissue of MRT, thus opening up a new era for the research of MRT origin. It is also concluded as follows: 1. Cellular tumorigenicity is different among differently杒aryotypiccells. 2. Highly variable strain of tumor cell line can be selected quickly and successfully in nude mouse. 3. Cellular tumorigenicity may be increased if chromosome aberration is very high. 4. The genetic characteristics of chromosomes of HeLa cells determines their tumorigenicity, chromosome number variation of HeLa cells has positive relationship with their carcinogenesis or tumorigenicity, and the turn of HeLa cells concerning their tumorigenicity from weak to strong is KB, X and NM20／X strains (excluding H strain, in which tumorigenicity remains to be determined by further experiments) respectively.